Treatment of asthma with MEK inhibitors

ABSTRACT

This invention provides a method of preventing or treating asthma by administering to a patient in need of treatment an effective amount of a selective MEK inhibitor, especially a phenyl amine of Formula I and II:

CROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application is a divisional application of U.S. applicationSer. No. 09/889,091 filed on Jul. 11, 2001, which is a 371 applicationof PCT/US99/30419 filed on Dec. 21, 1999, which claims the benefit ofpriority to U.S. Provisional Application Serial No. 60/115086 filed onJan. 7, 1999.

FIELD OF THE INVENTION

[0002] This invention relates to a method for preventing and treatingasthma in mammals comprising administering a compound characterized asan inhibitor of a family of enzymes known as MEK kinases, which aregroups of MAP (mitogen-associated protein kinase) and Erk (extracellularsignal-regulated) Kinases. These are enzymes that regulatephosphorylation of substrates in mammals.

BACKGROUND OF THE INVENTION

[0003] Asthma is a heterogeneous disorder of the airways that afflictsmillions of people. Airway inflammation, hyperresponsiveness, andobstruction characterize the condition. The disease often causes spasmsof the bronchial smooth muscle system, and affects both the upper andlower respiratory tracts. There are several forms of asthma,characterized by varying degrees of severity. Mild asthma, for example,is defined as brief episodes of wheezing, with or without dyspnea orcough. Moderately severe asthma is defined as wheezing and dyspnea, andcan be with or without cough and expectoration, but generally interfereswith daily activities and/or sleeping. Severe asthma is characterized byincapacitation due to dyspnea, and the afflicted patient typically isunable to eat or sleep normally, is very anxious, and is oftenexhausted. A condition known as status asthmaticus is the most severeform of asthma, and generally requires intensive hospital care, and mayeven prove fatal. The disease may occur as a result of both allergic andnonallergic mechanisms.

[0004] While there are several treatments available for relieving thesymptoms and discomfort associated with asthma, there are no cures.Moreover, the current treatments often cause side effects thatexacerbate the discomfort and precipitate other debilitating conditions.Mild asthma generally is treated with beta-adrenergic drugs, as well asantihistamines, especially in the case of children, to prevent or abortsporadic episodes. Moderately severe and severe asthma are generallytreated with adrenergic agents and bronchodilators, as well ascorticosteroids. Other actions caused by antiasthmatic agents whichlimit their widespread use include headache, fatigue, dry mouth,nervousness, and in some cases addiction and substance abuse. Recentadvances in the understanding of the pathogenesis and treatment ofasthma is discussed more fully by Grayson et al., The Mount SinaiJournal of Medicine, September 1998;65(4):246-256.

[0005] Because asthma is so prevalent in both children and adults, thereis an on-going need for agents that can treat the disease, or at leastrelieve the symptoms that accompany the disease, without causingundesirable side effects. We have now discovered that MEK inhibitors areparticularly useful for treating asthma and relieving the symptoms thataccompany the disease. An object of this invention is therefore toprovide a new method for preventing and treating asthmatic conditions.

SUMMARY OF THE INVENTION

[0006] This invention provides a method of preventing and treatingasthma, said method comprising the step of administering to a patient anantiasthmatic-effective amount of a MEK inhibitor. Selective MEKinhibitors are those compounds which inhibit the MEK 1 and MEK 2 enzymeswithout substantial inhibition of other such enzymes. In a preferredembodiment, the invention provides a method for preventing or treatingasthma by administering a MEK inhibitor. In a further embodiment, theinvention provides a method for preventing and/or treating asthmacomprising administering an effective amount of the selective MEKinhibitor described in U.S. Pat. No. 5,525,625, incorporated herein byreference, which selective MEK inhibitor is2-(2-amino-3-methoxyphenyl)-4-oxo-4H-[1]benzopyran.

[0007] In another preferred embodiment, the MEK inhibitor to beadministered is a phenyl amine derivative of Formula I:

[0008] In Formula (I), R₁ is hydrogen, hydroxy, C₁-C₈ alkyl, C₁-C₈alkoxy, halo, trifluoromethyl, or CN. R₂ is hydrogen. R₃, R₄, and R₅ areindependently selected from hydrogen, hydroxy, halo, trifluoromethyl,C₁-C₈ alkyl, C₁-C₈ alkoxy, nitro, CN, and —(O or NH)_(m)—(CH₂)_(n—R) ₉.R₉ is hydrogen, hydroxy, COOH, or NR₁₀R₁₁; n is 0-4; m is 0 or 1. Eachof R₁₀ and R₁₁ is independently selected from hydrogen and C₁-C₈ alkyl,or taken together with the nitrogen to which they are attached cancomplete a 3-10 member cyclic ring optionally containing 1, 2, or 3additional heteroatoms selected from O, S, NH, or N—(C₁-C₈ alkyl). Z isCOOR₇, tetrazolyl, CONR₆R₇, CONHNR₁₀R₁₁, or CH₂OR₇. R₆ and R₇independently are hydrogen, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl,(CO)—C₁-C₈ alkyl, aryl, heteroaryl, C₃-C₁₀ cycloalkyl, or C₃-C₁₀(cycloalkyl optionally containing one, two, or three heteroatomsselected from O, S, NH, or N alkyl); or R₆ and R₇ together with thenitrogen to which they are attached complete a 3-10 member cyclic ringoptionally containing 1, 2, or 3 additional heteroatoms selected from O,S, NH, or N alkyl. In formula (I), any of the foregoing alkyl, alkenyl,aryl, heteroaryl, heterocyclic, and alkynyl groups can be unsubstitutedor substituted by halo, hydroxy, C₁-C₆ alkoxy, amino, nitro, C₁-C₄alkylamino, di(C₁-C₄)alkylamino, C₃-C₆ cycloalkyl, phenyl, phenoxy,C₃-C₅ heteroaryl or heterocyclic radical, or C₃-C₅ heteroaryloxy orheterocyclic radical-oxy. The invention also provides a pharmaceuticallyacceptable salt, ester, amide, or prodrug of each of the disclosed MEKinhibitors.

[0009] Preferred embodiments of Formula (I) have a structure wherein:(a) R₁ is hydrogen, methyl, methoxy, fluoro, chloro, or bromo; (b) R₂ ishydrogen; (c) R₃, R₄, and R₅ independently are hydrogen, fluoro, chloro,bromo, iodo, methyl, methoxy, or nitro; (d) R₁₀ and R₁₁ independentlyare hydrogen or methyl; (e) Z is COOR₇, tetrazolyl, CONR₆R₇,CONHNR₁₀R₁₁, or CH₂OR₇; R₆ and R₇ independently are hydrogen, C₁₋₄alkyl, heteroaryl, or C₃₋₅ cycloalkyl optionally containing one or twoheteroatoms selected from O, S, or NH; or R₆ and R₇ together with thenitrogen to which they are attached complete a 5-6 member cyclic ringoptionally containing 1 or 2 additional heteroatoms selected from O, NHor N-alkyl; and wherein any of the foregoing alkyl or aryl groups can beunsubstituted or substituted by halo, hydroxy, methoxy, ethoxy, orheteroaryloxy (such as 2,3,4,5,6-pentafluorophenyl); (f) Z is COOR₇; (g)R₇ is H, pentafluorophenyl, or tetrazolyl; (h) R₃, R₄, and R₅ areindependently H, fluoro, or chloro; (i) R₄ is fluoro; (j) two of R₃, R₄,and R₅ are fluoro; or (k) or combinations of the above. In anotherpreferred embodiment of Formula (I), R₁ is methyl, fluoro, chloro, orbromo.

[0010] In a more preferred embodiment, the MEK inhibitor is selectedfrom a compound in Formula (I) Compound Table below.

Formula (I) Compound Table (Page 1 of 10)

[0011][4-Chloro-2-(1H-tetrazol-5-yl)-phenyl-(4-iodo-2-methyl-phenyl)-amine

[0012] (4-iodo-2-methyl-phenyl)-[2-(1H-tetrazol-5-yl)-phenyl]amine

[0013][4-nitro-2-(1H-tetrazol-5-yl)-phenyl-(4-iodo-2-methyl-phenyl)-amine

[0014] 4-Fluoro-2-(4-iodo-2-methylphenylamino)benzoic acid

[0015] 3,4,5-Trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0016] 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0017] 5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0018] 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0019] Sodium 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoate

[0020] 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0021] 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-benzoic acid

[0022] 4-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0023] 2-(4-Iodo-2-methyl-phenylamino)-benzoic acid

[0024] 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0025] 5-Iodo-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0026] 2,3,5-Trifluoro-4-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0027] 2-(4-Iodo-phenylamino)-5-methoxy-benzoic acid

[0028] 5-Methyl-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0029] 2-(4-Iodo-2-methyl-phenylamino)-4-nitro-benzoic acid

[0030] 2-(4-Bromo-2-methyl-phenylamino)-4-fluoro-benzoic acid

[0031] 2-(2-Bromo-4-iodo-phenylamino)-5-nitro-benzoic acid

[0032] 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-benzoic acid

[0033]5-Chloro-N-(2-hydroxyethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0034] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0035] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-benzamide

[0036] N-Ethyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0037] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N,N-dimethyl-benzamide

[0038]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(1H-tetrazol-5-yl)-benzamide

Formula (I) Compound Table (Continued, Page 2 of 10)

[0039] 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0040] 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N,N-dimethyl-benzamide

[0041] [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoylamino]-aceticacid

[0042] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-propyl-benzamide

[0043]5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0044] N,N-Diethyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0045]4-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0046] N,N-Diethyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0047] N-Butyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0048] 5-Chloro-N,N-diethyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0049] 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N,N-dimethyl-benzamide

[0050]5-Bromo-3,4-difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0051]N-(2,3-Dihydroxy-propyl)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0052]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl-ethyl)-benzamide

[0053]3,4-Difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0054]N-(2,3-Dihydroxy-propyl)4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0055]3,4-Difluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0056]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0057]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin4-yl-ethyl)-benzamide

Formula (I) Compound Table (Continued, Page 3 of 10)

[0058]4-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0059]5-Bromo-N-(3-dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-2-methylphenylamino)-benzamide

[0060]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0061]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0062]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0063]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-4-yl-ethyl)-benzamide

[0064]N-(3-Dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0065] N-Benzyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0066]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-hydroxy-ethyl)-benzamide

[0067]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0068]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl-propyl)-benzamide

[0069]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl-propyl)-benzamide

[0070]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-thiophen-2-yl-ethyl)-benzamide

[0071]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0072]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-morpholin4-yl-ethyl)-benzamide

Formula (I) Compound Table (Continued, Page 4 of 10)

[0073]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl-benzamide

[0074]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl-benzamide

[0075]2-(4-Bromo-2-methyl-phenylamino)-N-(3-dimethylamino-propyl)-3,4-difluoro-benzamide

[0076]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl-enzamide

[0077]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-4-yl-ethyl)-benzamide

[0078]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-pyridin-4-yl-ethyl)-benzamide

[0079]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(3-hydroxy-propyl)-benzamide

[0080]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0081] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-phenethyl-benzamide

[0082]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-thiophen-2-yl-ethyl)-benzamide

[0083]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-pyridin-4-ylmethyl-benzamide

[0084]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-phenethyl-benzamide

[0085]2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-piperidin-1-yl-ethyl)-benzamide

[0086]5-Chloro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0087]5-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0088] 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-pyridin-4-ylmethyl-benzamide

Formula (I) Compound Table (Continued, Page 5 of 10)

[0089] 5-Bromo-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl-propyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0090]5-Chloro-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0091]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl-ethyl)-benzamide

[0092](3-Hydroxy-pyrrolidin-1-yl)-[5-nitro-2-(4-iodo-2-methyl-phenylamino)-phenyl}-methanone

[0093]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0094]5-Bromo-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0095]N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0096]N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0097]N-{3-[4-(2-Hydroxy-ethyl)-piperazin-1-yl]-propyl}-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0098]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl-benzamide

[0099]5-Bromo-2-(4-iodo-2-ethyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0100]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl-ethyl)-benzamide

[0101]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-benzamide

[0102]5-Chloro-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0103]N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

Formula (I) Compound Table (Continued, Page 6 of 10)

[0104]5-Chloro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0105]5-Chloro-N-(3-diethylamino-2-hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0106]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl-ethyl)-benzamide

[0107]5-Bromo-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0108]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl-propyl)-benzamide

[0109]N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0110]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0111]5-Chloro-N-(3-diethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0112]5-Chloro-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0113]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl-propyl)-benzamide

[0114]2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(2-piperidin-1-yl-ethyl)-benzamide

[0115]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperazin-1-yl-ethyl)-benzamide

[0116]N-(2-Diethylamino-ethyl)-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0117]5-Bromo-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0118]N-(3-Hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

Formula (I) Compound Table (Continued, Page 7 of 10)

[0119]5-Fluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0120]N-(3-Diethylamino-propyl)-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0121]N-(3-Diethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0122]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0123]2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(3-piperidin-1-yl-propyl)-benzamide

[0124] [5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-(2 or3-hydroxy-pyrrolidin-1-yl)-methanone

[0125]5-Bromo-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0126]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl-ethyl)-benzamide

[0127]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl-propyl)-benzamide

[0128][5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-4-(2-hydroxy-ethyl)-piperazin-1-yl)-methanone

[0129]N-(3-Diethylamino-2-hydroxy-propyl)-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0130] N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0131]5-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0132]5-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0133] N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0134] N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

Formula (I) Compound Table (Continued, Page 8 of 10)

[0135]2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(4-sulfamoyl-benzyl)-benzamide

[0136]5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0137]N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0138]N-(2-Hydroxy-ethyl)-2-(4-iodo-2-ethyl-phenylamino)-5-nitro-benzamide

[0139]2-(4-Iodo-2-methyl-phenylamino)-N-methyl-5-nitro-N-phenyl-benzamide

[0140] 5-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0141]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

[0142] N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0143] N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0144]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)-benzamide

[0145] N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0146] N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0147] 5-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0148]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

[0149]5-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)-benzamide

[0150]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)-benzamide

[0151] N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0152]2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(4-sulfamoyl-benzyl)-benzamide

[0153] N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0154]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-benzamide

[0155] N-Cyclopropyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

Formula (I) Compound Table (Continued, Page 9 of 10)

[0156]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

[0157] N-Benzyloxy-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0158] N-Cyclohexyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0159] N-Allyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0160]5-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-benzamide

[0161]2-(4-Iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-5-nitro-benzamide

[0162]5-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

[0163] N-Cyclohexyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0164] 5-Chloro-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0165]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-benzamide

[0166] 5-Bromo-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0167]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-benzamide

[0168] N-Cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0169] N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0170] N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0171]5-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0172]5-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0173]2-(4-Iodo-2-methyl-phenylamino)-N-methyl-5-nitro-N-phenyl-benzamide

[0174]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

[0175]N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0176] 5-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0177] N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0178]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-benzamide

Formula (I) Compound Table (Continued, Page 10 of 10)

[0179]N-(2-Hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0180]5-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0181] 5-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0182] N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0183]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)-benzamide

[0184] N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0185] N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0186] N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0187] N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0188]5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)-benzamide

[0189] N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide

[0190] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzyl alcohol

[0191] [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-methanol

[0192] [2-(4-Iodo-2-methyl-phenylamino)-5-nitro-phenyl]-methanol

[0193] [5-Bromo-2-(4-iodo-2-methyl-phenylamino)-phenyl]-methanol

[0194] N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide.

[0195] In another preferred embodiment, the MEK inhibitor is a compoundof Formula II

[0196] In Formula (II), R_(1a) is hydrogen, hydroxy, C₁-C₈ alkyl, C₁-C₈alkoxy, halo, trifluoromethyl, or CN. R_(2a) is hydrogen. Each ofR_(3a), R_(4a), and R_(5a) is independently selected from hydrogen,hydroxy, halo, trifluoromethyl, C₁-C₈ alkyl, C₁-C₈ alkoxy, nitro, CN,and (O or NH)_(m)—(CH₂)_(n)—R_(9a). R_(9a) is hydrogen, hydroxy, CO₂H orNR_(10a)R_(11a); n is 0-4; and m is 0 or 1. Each of R_(10a) and R_(11a)is independently hydrogen or C₁-C₈ alkyl, or taken together with thenitrogen to which they are attached can complete a 3- to 10-membercyclic ring optionally containing one, two, or three additionalheteroatoms selected from O, S, NH, or N—(C₁-C₈ alkyl). R_(6a) ishydrogen, C₁-C₈ alkyl, (CO)—(C₁-C₈ alkyl), aryl, aralkyl, or C₃-C₁₀cycloalkyl. R_(7a) is hydrogen, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈alkynyl, C₃-C₁₀ (cycloalkyl or cycloalkyl optionally containing aheteroatom selected from O, S, or NR_(9a)). In Formula (II), any of theforegoingany of the foregoing alkyl, alkenyl, aryl, heteroaryl,heterocyclic, and alkynyl groups can be unsubstituted or substituted byhalo, hydroxy, C₁-C₆ alkoxy, amino, nitro, C₁-C₄ alkylamino,di(C₁-C₄)alkylamino, C₃-C₆ cycloalkyl, phenyl, phenoxy, C₃-C₅ heteroarylor heterocyclic radical, or C₃-C₅ heteroaryloxy or heterocyclicradical-oxy; or R_(6a) and R_(7a) taken together with the N to whichthey are attached can complete a 5- to 10-membered cyclic ring,optionally containing one, two, or three additional heteroatoms selectedfrom O, S, or NR_(10a)R_(11a). The invention also encompassespharmaceutically acceptable salts, esters, amides or prodrugs of each ofthe disclosed compounds.

[0197] Preferred embodiments of Formula (II) are those structureswherein: (a) R_(1a) is H, methyl, fluoro, or chloro; (b) R_(2a) is H;R_(3a), R_(4a) and R_(5a) are each H, Cl, nitro, or F; (c) R_(6a) is H;(d) R_(7a) is methyl, ethyl, 2-propenyl, propyl, butyl, pentyl, hexyl,cyclopropylmethyl, cyclobutyl methyl, cyclopropylmethyl, orcyclopropylethyl; (e) the 4′ position is I, rather than Br; (f) R_(4a)is F at the 4 position, para to the CO—N—R_(6a)—OR_(7a) group and metato the bridging nitrogen; (f) R_(3a) or R_(5a) is F; (g) at least one ofR_(3a), R_(4a) and R_(5a) is F; (h) R_(1a) is methyl or chloro; or (i)or a combination of the above.

[0198] In a more preferred embodiment the MEK inhibitor is a compoundselected from Formula (II) Compound Table below.

Formula (II) Compound Table (Page 1 of 7)

[0199] 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0200] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(methoxy)-benzamide

[0201]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(prop-2-ynyloxy)-benzamide

[0202]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-phenoxyethoxy)-benzamide

[0203]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-thienylmethoxy)-benzamide

[0204]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(prop-2-enyloxy)-benzamide

[0205]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclopropylmethoxy)-benzamide

[0206]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclopentoxy)-benzamide

[0207]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-furylmethoxy)-benzamide

[0208] 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-ethoxy-benzamide

[0209]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(but-2-enyloxy)-benzamide

[0210]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclopropylmethoxy)-benzamide

[0211]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(1-methylprop-2-ynyloxy)-benzamide

[0212]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-phenylprop-2-ynyloxy)-benzamide

[0213]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-5-phenylpent-2-en-4-ynyloxy)-benzamide

[0214]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(prop-2-ynyloxy)-benzamide

[0215]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(propoxy)-benzamide

Formula (II) Compound Table (Continued, Page 2 of 7)

[0216]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclobutyloxy)-benzamide

[0217]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-thienylmethoxy)-benzamide

[0218]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-methyl-prop-2-enyloxy)-benzamide

[0219]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-phenoxyethoxy)-benzamide

[0220]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(but-2-enyloxy)-benzamide

[0221]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(but-3-ynyloxy)-benzamide

[0222]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclopentyloxy)-benzamide

[0223]3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-(2-fluorophenyl)-prop-2-ynyloxy)-benzamide

[0224]5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0225]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(n-propoxy)-benzamide

[0226]5-Bromo-3,4-difluoro-N-(furan-3-ylmethoxy)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0227]5-Bromo-N-(but-2-enyloxy)-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0228]5-Bromo-N-butoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0229]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-but-2-enyloxy)-benzamide

[0230]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-pent-2-en-4-ynyloxy)-benzamide

Formula (II) Compound Table (Continued, Page 3 of 7)

[0231]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-benzyl)-N-[5-(3-methoxy-phenyl)-3-methyl-pent-2-en-4-ynyloxy]-benzamide

[0232]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(prop-2-ynyloxy)-benzamide

[0233]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-[3-(3-methoxy-phenyl)-prop-2-ynyloxy]-benzamide

[0234]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(thiopen-2-ylmethoxy)-benzamide

[0235]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(pyridin-3-ylmethoxy)-benzamide

[0236]5-Bromo-3-4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-(2-fluorophenyl)-prop-2-ynyloxy)-benzamide

[0237]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(ethoxy)-benzamide

[0238]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(cyclopropylmethoxy)-benzamide

[0239]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(isopropoxy)-benzamide

[0240]5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-but-3-ynyloxy)-benzamide

[0241] 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0242]5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(tetrahydro-pyran-2-yloxy)-benzamide

[0243] 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methoxy-benzamide

[0244] 4-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-phenylmethoxy-benzamide

[0245]4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-phenylmethoxy-benzamide

[0246] 5-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0247] 5-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-phenylmethoxy-benzamide

[0248]5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(tetrahydropyran-2-yloxy)-benzamide

Formula (II) Compound Table (Continued, Page 4 of 7)

[0249]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(3-phenylprop-2-ynyloxy)-benzamide

[0250]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(3-furylmethoxy)-benzamide

[0251]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(2-thienylmethoxy)-benzamide

[0252]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(but-3-ynyloxy)-benzamide

[0253]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(2-methyl-prop-2-enyloxy)-benzamide

[0254]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(but-2-enyloxy)-benzamide

[0255]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(methoxy)-benzamide

[0256]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(ethoxy)-benzamide

[0257]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(cyclobutoxy)-benzamide

[0258]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(isopropoxy)-benzamide

[0259]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(2-phenoxyethoxy)-benzamide

[0260]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(cyclopropylmethoxy)-benzamide

[0261]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(n-propoxy)-benzamide

[0262]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(1-methyl-prop-2-ynyloxy)-benzamide

[0263]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(3-(3-fluorophenyl)-prop-2-ynyloxy)-benzamide

[0264]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(4,4-dimethylpent-2-ynyloxy)-benzamide

Formula (II) Compound Table (Continued, Page 5 of 7)

[0265]3,4-Difluoro-2-(4-bromo-2-methyl-phenylamino)-N-(cyclopentoxy)-benzamide

[0266]3,4,5-Trifluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0267]5-Chloro-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0268]5-Bromo-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide

[0269] N-Hydroxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzamide

[0270]3,4,5-Trifluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide

[0271]5-Chloro-3,4-difluoro-2-(2-fluoro4-iodo-phenylamino)-N-hydroxy-benzamide

[0272]5-Bromo-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide

[0273] 2-(2-Fluoro-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide

[0274]2-(2-Chloro-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide

[0275]5-Chloro-2-(2-chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide

[0276]5-Bromo-2-(2-bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide

[0277] 2-(2-Chloro-4-iodo-phenylamino)-N-hydroxy-4-methyl-benzamide

[0278]2-(2-Bromo-4-iodo-phenylamino)-3,4,5-trifluoro-N-hydroxy-benzamide

[0279]2-(2-Bromo-4-iodo-phenylamino)-5-chloro-3,4-difluoro-N-hydroxy-benzamide

[0280] 2-(2-Bromo-4-iodo-phenylamino)-N-hydroxy-4-nitro-benzamide

[0281] 4-Fluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide

[0282] 3,4-Difluoro-2-(2-fluoro-4-iodo-phenylamino)-N-hydroxy-benzamide

[0283] 2-(2-Chloro4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide

[0284] 2-(2-Chloro-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide

[0285] 2-(2-Bromo-4-iodo-phenylamino)-4-fluoro-N-hydroxy-benzamide

[0286] 2-(2-Bromo-4-iodo-phenylamino)-3,4-difluoro-N-hydroxy-benzamide

Formula (II) Compound Table (Continued, Page 6 of 7)

[0287]N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0288]5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0289]5-Bromo-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide

[0290]N-Cyclopropylmethoxy-2-(4-iodo-2-methyl-phenylamino)-4-nitro-benzamide

[0291]N-Cyclopropylmethoxy-3,4,5-trifluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide

[0292]5-Chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide

[0293]5-Bromo-2-(2-chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide

[0294]N-Cyclopropylmethoxy-2-(2-fluoro-4-iodo-phenylamino)-4-nitro-benzamide

[0295]2-(2-Chloro-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide

[0296]5-Chloro-2-(2-chloro4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide

[0297]5-Bromo-2-(2-bromo4-iodo-phenylamino)-N-ethoxy-3,4-difluoro-benzamide

[0298] 2-(2-Chloro-4-iodo-phenylamino)-N-ethoxy-4-nitro-benzamide

[0299]2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4,5-trifluoro-benzamide

[0300]2-(2-Bromo-4-iodo-phenylamino)-5-chloro-N-cyclopropylmethoxy-3,4-difluoro-benzamide

[0301]2-(2-Bromo4-iodo-phenylamino)-N-cyclopropylmethoxy-4-nitro-benzamide

Formula (II) Compound Table (Continued, Page 7 of 7)

[0302]N-Cyclopropylmethoxy-4-fluoro-2-(2-fluoro4-iodo-phenylamino)-benzamide

[0303]N-Cyclopropylmethoxy-3,4-difluoro-2-(2-fluoro-4-iodo-phenylamino)-benzamide

[0304]2-(2-Chloro4-iodo-phenylamino)-N-cyclopropylmethoxy-4-fluoro-benzamide

[0305]2-(2-Chloro4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide

[0306]2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy4-fluoro-benzamide

[0307]2-(2-Bromo-4-iodo-phenylamino)-N-cyclopropylmethoxy-3,4-difluoro-benzamide.

[0308] In the most preferred embodiment of this invention, a compoundselected from the following is administered to a patient (ie, a mammal)in an amount that is effective to prevent or treat rheumatoid arthritisor osteoarthritis:

[0309]2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide(PD184352); 2-(2-Methyl4-iodophenylamino)-N-hydroxy-4-fluorobenzamide(PD170611);2-(2-Methyl-4-iodophenylamino)-N-hydroxy-3,4-difluoro-5-bromobenzamide(PD171984);2-(2-Methyl-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-bromobenzamide(PD177168);2-(2-Methyl-4-iodophenylamino)-N-cyclobutylmethoxy-3,4-difluoro-5-bromobenzamide(PD 180841);2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-bromobenzamide(PD 184161);2-(2-Chloro-4-iodophenylamino)-N-hydroxy-3,4-difluoro-5-bromobenzamide(PD184386);2-(2-Chloro-4-iodophenylamino)-N-cyclobutylmethoxy-3,4-difluorobenzamide(PD 185625); 2-(2-Chloro-4-iodophenylamino)-N-hydroxy-4-fluorobenzamide(PD 185848);2-(2-Methyl-4-iodophenylamino)-N-hydroxy-3,4-difluorobenzamide (PD188563);2-(2-Methyl4-iodophenylamino)-N-cyclopropylmethoxy-3,4,5-trifluorobenzamide(PD 198306); and2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-4-fluorobenzamide(PD 203311); and the benzoic acid derivatives thereof. For example, thebenzoic acid derivative of PD 198306 is2-(2-Methyl-4-iodophenylamino)-3,4,5-trifluorobenzoic acid.

[0310] Additional preferred compounds include2-(2-chloro-4-iodophenylamino)-5-chloro-N-cyclopropylmethoxy-3,4-difluorobenzamide(PD 297189),2-(4-iodophenylamino)-N-cyclopropylmethoxy-5-chloro-3,4-difluorobenzamide(PD 297190), 2-(4-iodophenylamino)-5-chloro-3,4-difluorobenzoic acid (PD296771), 2-(2-chloro4-iodophenylamino)-5-chloro-3,4-difluorobenzoic acid(PD 296770),5-chloro-3,4-difluoro-2-(4-iodo-2-methylphenylamino)-benzoic acid (PD296767); and5-chloro-N-cyclopropylmethoxy-3,4-difluoro-2-(4-iodo-2-methylphenylamino)-benzamide(PD 298127).

[0311] The invention further provides methods of synthesis and syntheticintermediates.

[0312] Other features and advantages of the invention are apparent fromthe detailed description, examples, and claims set forth.

[0313] In a further preferred embodiment of this invention, a mitoticinhibitor is administered to a patient suffering from cancer and in needof treatment in combination with a selective MEK inhibitor selectedfrom:2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluorobenzamide(PD184352); 2-(2-Methyl-4-iodophenylamino)-N-hydroxy-4-fluorobenzamide(PD170611);2-(2-Methyl-4-iodophenylamino)-N-hydroxy-3,4-difluoro-5-bromobenzamide(PD171984), a more preferred compound;2-(2-Methyl-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-bromobenzamide(PD177168);2-(2-Methyl-4-iodophenylamino)-N-cyclobutylmethoxy-3,4-difluoro-5-bromobenzamide(PD 180841);2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-3,4-difluoro-5-bromobenzamide(PD 184161);2-(2-Chloro-4-iodophenylamino)-N-hydroxy-3,4-difluoro-5-bromobenzamide(PD184386);2-(2-Chloro-4-iodophenylamino)-N-cyclobutylmethoxy-3,4-difluorobenzamide(PD 185625); 2-(2-Chloro4-iodophenylamino)-N-hydroxy-4-fluorobenzamide(PD 185848);2-(2-Methyl4-iodophenylamino)-N-hydroxy-3,4-difluorobenzamide(PD188563);2-(2-Methyl-4-iodophenylamino)-N-cyclopropylmethoxy-3,4,5-trifluorobenzamide(PD 198306); and2-(2-Chloro-4-iodophenylamino)-N-cyclopropylmethoxy-4-fluorobenzamide(PD 203311); and the benzoic acid derivatives thereof. For example, thebenzoic acid derivative of PD 198306 is2-(2-Methyl-4-iodophenylamino)-3,4,5-trifluorobenzoic acid.

DETAILED DESCRIPTION OF THE INVENTION

[0314] This invention provides a method of preventing or treating asthmain a patient which comprises administering to a patient suffering fromasthma and in need of treatment, or to a patient at risk for developingan asthmatic attack, an anti-asthmatic effective amount of a MEKinhibitor. The invention provides a method of preventing and treatingall forms of asthma and relieving the symptoms that accompany thedisease. The invention is preferably practiced by administering a phenylamine MEK inhibitor of Formula I or Formula II. Such MEK phenyl aminecompounds are specific MEK 1 and MEK 2 inhibitors, meaning that theyinhibit these enzymes without inhibiting other enzymes to a greatextent.

[0315] The compounds of the present invention, which can be used totreat septic shock, are MEK inhibitors. A MEK inhibitor is a compoundthat shows MEK inhibition when tested in the assays titled “EnzymeAssays” in U.S. Pat. No. 5,525,625, column 6, beginning at line 35. Thecomplete disclosure of U.S. Pat. No. 5,525,625 is hereby incorporated byreference. An example of a MEK inhibitor is2-(2-amino-3-methoxyphenyl)-4-oxo-4H-[1]benzopyran. Specifically, acompound is a MEK inhibitor if a compound shows activity in the assaytitled “Cascade Assay for Inhibitors of the MAP Kinase Pathway,” column6, line 36 to column 7, line 4 of the U.S. Pat. No. 5,525,625 and/orshows activity in the assay titled “In Vitro MEK Assay” at column 7,lines 4 to 27 of the above-referenced patent.

[0316] A. Terms

[0317] Some of the terms used herein are defined below and by theirusage throughout this disclosure.

[0318] The term “patient” means all animals including humans. Examplesof patients include humans, cows, dogs, cats, goats, sheep, horses, andpigs. The mammals to be treated according to this invention are patientswho have developed asthma and are suffering from the symptoms associatedwith disease, or who are at risk for developing the disease, for examplehaving a family history of asthma. Those skilled in the medical art arereadily able to identify individual patients, particularly children, whoare afflicted with asthma, as well as those who are susceptible todeveloping the disease.

[0319] As used herein, the term “aryl” means a cyclic, bicyclic, ortricyclic aromatic ring moiety having from five to twelve carbon atoms.Examples of typical aryl groups include phenyl, naphthyl, and fluorenyl.The aryl may be substituted by one, two, or three groups selected fromfluoro, chloro, bromo, iodo, alkyl, hydroxy, alkoxy, nitro, amino,alkylamino, or dialkylamino. Typical substituted aryl groups include3-fluorophenyl, 3,5-dimethoxyphenyl, 4-nitronaphthyl,2-methyl-4-chloro-7-aminofluorenyl, and the like.

[0320] The term “aryloxy” means an aryl group bonded through an oxygenatom, for example phenoxy, 3-bromophenoxy, naphthyloxy, and4-methyl-1-fluorenyloxy.

[0321] “Heteroaryl” means a cyclic, bicyclic, or tricyclic aromatic ringmoiety having from four to eleven carbon atoms and one, two, or threeheteroatoms selected from O, S, or N. Examples include furyl, thienyl,pyrrolyl, pyrazolyl, imidazolyl, triazolyl, thiazolyl, oxazolyl,xanthenyl, pyronyl, indolyl, pyrimidyl, naphthyridyl, pyridyl,benzinnidazolyl, and triazinyl. The heteroaryl groups can beunsubstituted or substituted by one, two, or three groups selected fromfluoro, chloro, bromo, iodo, alkyl, hydroxy, alkoxy, nitro, amino,alkylamino, or dialkylamino. Examples of substituted heteroaryl groupsinclude chloropyranyl, methylthienyl, fluoropyridyl,amino-1,4-benzisoxazinyl, nitroisoquinolinyl, and hydroxyindolyl.

[0322] The heteroaryl groups can be bonded through oxygen to makeheteroaryloxy groups, for example thienyloxy, isothiazolyloxy,benzofuranyloxy, pyridyloxy, and 4-methylisoquinolinyloxy.

[0323] The term “alkyl” means straight and branched chain aliphaticgroups. Typical alkyl groups include methyl, ethyl, isopropyl,tert.-butyl, 2,3-dimethylhexyl, and 1,1-dimethylpentyl. The alkyl groupscan be unsubstituted or substituted by halo, hydroxy, alkoxy, amino,alkylamino, dialkylamino, cycloalkyl, aryl, aryloxy, heteroaryl, orheteroaryloxy, as those terms are defined herein. Typical substitutedalkyl groups include chloromethyl, 3-hydroxypropyl,2-dimethylaminobutyl, and 2-(hydroxymethylamino)ethyl. Examples of aryland aryloxy substituted alkyl groups include phenylmethyl,2-phenylethyl, 3-chlorophenylmethyl,1,1-dimethyl-3-(2-nitrophenoxy)butyl, and 3,4,5-trifluoronaphthylmethyl.Examples of alkyl groups substituted by a heteroaryl or heteroaryloxygroup include thienylmethyl, 2-furylethyl, 6-furyloxyoctyl,4-methylquinolyloxymethyl, and 6-isothiazolylhexyl. Cycloalkylsubstituted alkyl groups include cyclopropylmethyl, 2-cyclohexyethyl,piperidyl-2-methyl, 2-(piperidin-1-yl)-ethyl, 3-(morpholin-4-yl)propyl.

[0324] “Alkenyl” means a straight or branched carbon chain having one ormore double bonds. Examples include but-2-enyl, 2-methyl-prop-2-enyl,1,1-dimethyl-hex-4-enyl, 3-ethyl-4-methyl-pent-2-enyl, and3-isopropyl-pent-4-enyl. The alkenyl groups can be substituted withhalo, hydroxy, alkoxy, amino, alkylamino, dialkylamino, aryl, aryloxy,heteroaryl, or heteroyloxy, for example 2-bromoethenyl,3-hydroxy-2-butenyl, 1-aminoethenyl, 3-phenylprop-2-enyl,6-thienyl-hex-2-enyl, 2-furyloxy-but-2-enyl, and4-naphthyloxy-hex-2-enyl.

[0325] “Alkynyl” means a straight or branched carbon chain having atleast one triple bond. Typical alkynyl groups include prop-2-ynyl,2-methyl-hex-5-ynyl, 3,4-dimethyl-hex-5-ynyl, and 2-ethyl-but-3-ynyl.The alkynyl groups can be substituted as the alkyl and alkenyl groups,for example, by aryl, aryloxy, heteroaryl, or heteroaryloxy, for example4-(2-fluorophenyl)-but-3-ynyl, 3-methyl-5-thienylpent-4-ynyl,3-phenoxy-hex-4-ynyl, and 2-furyloxy-3-methyl-hex-4-ynyl.

[0326] The alkenyl and alkynyl groups can have one or more double bondsor triple bonds, respectively, or a combination of double and triplebonds. For example, typical groups having both double and triple bondsinclude hex-2-en-4-ynyl, 3-methyl-5-phenylpent-2-en-4-ynyl, and3-thienyloxy-hex-3-en-5-ynyl.

[0327] The term “cycloalkyl” means a nonaromatic ring or fused rings.Examples include cyclopropyl, cyclobutyl, cyclopenyl, cyclooctyl,bicycloheptyl, adamantyl, and cyclohexyl. The ring can optionallycontain one, two, or three heteroatoms selected from O, S, or N. Suchgroups include tetrahydrofuryl, tetrahydropyrrolyl,octahydrobenzofuranyl, morpholinyl, piperazinyl, pyrrolidinyl,piperidinyl, octahydroindolyl, and octahydrobenzothiofuranyl. Thecycloalkyl groups can be substituted with the same substituents as analkyl and alkenyl groups, for example, halo, hydroxy, aryl, andheteroaryloxy. Examples include 3-hydroxycyclohexyl, 2-aminocyclopropyl,2-phenylpyrrolidinyl, and 3-thienylmorpholine-1-yl.

[0328] Selective MEK 1 or MEK 2 inhibitors are those compounds whichinhibit the MEK 1 or MEK 2 enzymes, respectively, without substantiallyinhibiting other enzymes such as MKK3, PKC, Cdk2A, phosphorylase kinase,EGF, and PDGF receptor kinases, and C-src. In general, a selective MEK 1or MEK 2 inhibitor has an IC₅₀ for MEK 1 or MEK 2 that is at leastone-fiftieth ({fraction (1/50)}) that of its IC₅₀ for one of theabove-named other enzymes. Preferably, a selective inhibitor has an IC₅₀that is at least {fraction (1/100)}, more preferably {fraction (1/500)},and even more preferably {fraction (1/1000)}, {fraction (1/5000)}, orless than that of its IC₅₀ or one or more of the above-named enzymes.

[0329] B. Administration and Formulation

[0330] The MEK inhibitors of the present method can be administered to apatient as part of a pharmaceutically acceptable composition. Thecompositions can be administered to humans and animals either orally,rectally, parenterally (intravenously, intramuscularly,orsubcutaneously), intracisternally, intravaginally, intraperitoneally,intravesically, locally (powders, ointments, or drops), or as a buccalor nasal spray.

[0331] Compositions suitable for parenteral injection may comprisephysiologically acceptable sterile aqueous or nonaqueous solutions,dispersions, suspensions or emulsions, and sterile powders forreconstitution into sterile injectable solutions or dispersions.Examples of suitable aqueous and nonaqueous carriers, diluents,solvents, or vehicles include water, ethanol, polyols (propyleneglycol,polyethyleneglycol, glycerol, and the like), suitable mixtures thereof,vegetable oils (such as olive oil), and injectable organic esters suchas ethyl oleate. Proper fluidity can be maintained, for example, by theuse of a coating such as lecithin, by the maintenance of the requiredparticle size in the case of dispersions and by the use of surfactants.

[0332] These compositions may also contain adjuvants such as preserving,wetting, emulsifying, and dispensing agents. Prevention of the action ofmicroorganisms can be ensured by various antibacterial and antifungalagents, for example, parabens, chlorobutanol, phenol, sorbic acid, andthe like. It may also be desirable to include isotonic agents, forexample sugars, sodium chloride, and the like. Prolonged absorption ofthe injectable pharmaceutical form can be brought about by the use ofagents delaying absorption, for example, aluminum monostearate andgelatin.

[0333] Solid dosage forms for oral administration include capsules,tablets, pills, powders, and granules. In such solid dosage forms, theactive compound is admixed with at least one inert customary excipient(or carrier) such as sodium citrate or dicalcium phosphate or (a)fillers or extenders, as for example, starches, lactose, sucrose,glucose, mannitol, and silicic acid, (b) binders, as for example,carboxymethylcellulose, alignates, gelatin, polyvinylpyrrolidone,sucrose, and acacia, (c) humectants, as for example, glycerol, (d)disintegrating agents, as for example, agar-agar, calcium carbonate,potato or tapioca starch, alginic acid, certain complex silicates, andsodium carbonate, (e) solution retarders, as for example paraffin, (f)absorption accelerators, as for example, quaternary ammonium compounds,(g) wetting agents, as for example, cetyl alcohol and glycerolmonostearate, (h) adsorbents, as for example, kaolin and bentonite, and(i) lubricants, as for example, talc, calcium stearate, magnesiumstearate, solid polyethylene glycols, sodium lauryl sulfate, or mixturesthereof. In the case of capsules, tablets, and pills, the dosage formsmay also comprise buffering agents.

[0334] Solid compositions of a similar type may also be employed asfillers in soft and hard-filled gelatin capsules using such excipientsas lactose or milk sugar as well as high molecular weightpolyethyleneglycols, and the like.

[0335] Solid dosage forms such as tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells, such as entericcoatings and others well-known in the art. They may contain opacifyingagents, and can also be of such composition that they release the activecompound or compounds in a certain part of the intestinal tract in adelayed manner. Examples of embedding compositions which can be used arepolymeric substances and waxes. The active compounds can also be inmicro-encapsulated form, if appropriate, with one or more of theabove-mentioned excipients.

[0336] Liquid dosage forms for oral administration includepharmaceutically acceptable emulsions, solutions, suspensions, syrups,and elixirs. In addition to the active compounds, the liquid dosageforms may contain inert diluents commonly used in the art, such as wateror other solvents, solubilizing agents and emulsifiers, as for example,ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzylalcohol, benzyl benzoate, propyleneglycol, 1,3-butyleneglycol,dimethylformamide, oils, in particular, cottonseed oil, groundnut oil,corn germ oil, olive oil, castor oil and sesame oil, glycerol,tetrahydrofurfuryl alcohol, polyethyleneglycols, and fatty acid estersof sorbitan or mixtures of these substances, and the like.

[0337] Besides such inert diluents, the composition can also includeadjuvants, such as wetting agents, emulsifying and suspending agents,sweetening, flavoring, and perfuming agents.

[0338] Suspensions, in addition to the active compounds, may containsuspending agents, as for example, ethoxylated isostearyl alcohols,polyoxyethylene sorbitol and sorbitan esters, microcrystallinecellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth,or mixtures of these substances, and the like.

[0339] Compositions for rectal administrations are preferablysuppositories which can be prepared by mixing the compounds of thepresent invention with suitable non-irritating excipients or carrierssuch as cocoa butter, polyethyleneglycol, or a suppository wax, whichare solid at ordinary temperatures but liquid at body temperature andtherefore, melt in the rectum or vaginal cavity and release the activecomponent.

[0340] Dosage forms for topical administration of a compound of thisinvention include ointments, powders, sprays, and inhalants. The activecomponent is admixed under sterile conditions with a physiologicallyacceptable carrier and any preservatives, buffers, or propellants as maybe required. Ophthalamic formulations, eye ointments, powders, andsolutions are also contemplated as being within the scope of thisinvention.

[0341] The compounds of the present method can be administered to apatient at dosage levels in the range of about 0.1 to about 1000 mg perday. For a normal human adult having a body weight of about 70 kg, adosage in the range of about 0.01 to about 100 mg per kg of body weightper day is preferable. The specific dosage used, however, can vary. Forexample, the dosage can depend on a numbers of factors including therequirements of the patient, the severity of the condition beingtreated, and the pharmacological activity of the compound being used.The determination of optimum dosages for a particular patient iswell-known to those skilled in the art.

[0342] The compounds of the present method can be administered aspharmaceutically acceptable salts, esters, amides, or prodrugs. The term“pharmaceutically acceptable salts, esters, amides, and prodrugs” asused herein refers to those carboxylate salts, amino acid additionsalts, esters, amides, and prodrugs of the compounds of the presentinvention which are, within the scope of sound medical judgment,suitable for contact with the tissues of patients without unduetoxicity, irritation, allergic response, and the like, commensurate witha reasonable benefit/risk ratio, and effective for their intended use,as well as the zwitterionic forms, where possible, of the compounds ofthe invention.

[0343] The term “salts” refers to the relatively non-toxic, inorganicand organic acid addition salts of compounds of the present invention.These salts can be prepared in situ during the final isolation andpurification of the compounds or by separately reacting the purifiedcompound in its free base form with a suitable organic or inorganic acidand isolating the salt thus formed. Representative salts include thehydrobromide, hydrochloride, sulfate, bisulfate, nitrate, acetate,oxalate, valerate, oleate, palmitate, stearate, laurate, borate,benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate,succinate, tartrate, naphthylate, mesylate, glucoheptonate,lactiobionate and laurylsulphonate salts, and the like. These mayinclude cations based on the alkali and alkaline earth metals, such assodium, lithium, potassium, calcium, magnesium and the like, as well asnontoxic ammonium, quaternary ammonium, and amine cations including, butnot limited to ammonium, tetramethylammonium, tetraethylammonium,methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine,and the like. (See, for example, S. M. Berge, et al., “PharmaceuticalSalts,” J. Pham. Sci., 1977;66:1-19 which is incorporated herein byreference.) Examples of pharmaceutically acceptable, non-toxic esters ofthe compounds of this invention include C₁-C₆ alkyl esters wherein thealkyl group is a straight or branched chain. Acceptable esters alsoinclude C₅-C₇ cycloalkyl esters as well as arylalkyl esters such as, butnot limited to benzyl. C₁-C₄ alkyl esters are preferred. Esters of thecompounds of the present invention may be prepared according toconventional methods.

[0344] Examples of pharmaceutically acceptable, non-toxic amides of thecompounds of this invention include amides derived from ammonia, primaryC₁-C₆ alkyl amines and secondary C₁-C₆ dialkyl amines wherein the alkylgroups are straight or branched chain. In the case of secondary aminesthe amine may also be in the form of a 5 or 6 membered heterocyclecontaining one nitrogen atom. Amides derived from ammonia, C₁-C₃ alkylprimary amines and C₁-C₂ dialkyl secondary amines are preferred. Amidesof the compounds of the invention may be prepared according toconventional methods.

[0345] The term “prodrug” refers to compounds that are rapidlytransformed in vivo to yield the parent compound of the above formula,for example, by hydrolysis in blood. A thorough discussion is providedin T. Higuchi and V. Stella, “Pro-drugs as Novel Delivery Systems,” Vol.14 of the A.C.S. Symposium Series, and in Bioreversible Carriers in DrugDesign, ed. Edward B. Roche, American Pharmaceutical Association andPergamon Press, 1987, both of which are incorporated herein byreference.

[0346] In addition, the compounds of the present method can exist inunsolvated as well as solvated forms with pharmaceutically acceptablesolvents such as water, ethanol, and the like. In general, the solvatedforms are considered equivalent to the unsolvated forms for the purposesof the present invention.

[0347] Some of the compounds of the present method can exist indifferent stereoisometric forms by virtue of the presence of chiralcenters. It is contemplated that all stereoisometric forms of thecompounds as well as mixtures thereof, including racemic mixtures, formpart of this invention.

[0348] C. Synthesis

[0349] The examples presented below are intended to illustrateparticular embodiments of the invention and are not intended to limitthe scope of the specification, including the claims, in any way. Afterthe priority date of the present disclosure, related syntheses and MEKinhibition data were also published in WO 99/01421 and WO 99/01426,hereby incorporated by reference.

[0350] The 2-(4-bromo and 4-iodo phenylamino)-benzoic acid derivativesof Formula I can be prepared from commercially available startingmaterials utilizing synthetic methodologies well-known to those skilledin organic chemistry. A typical synthesis is carried out by reacting a4-bromo or 4-iodo aniline with a benzoic acid having a leaving group atthe 2-position to give a 2-(phenylamino)-benzoic acid. This process isdepicted in Scheme 1.

[0351] where L is a leaving group, for example halo such as fluoro.

[0352] The reaction of aniline and the benzoic acid derivative generallyis accomplished by mixing the benzoic acid with an equimolar quantity orexcess of the aniline in an unreactive organic solvent such astetrahydrofuran or toluene, in the presence of a base such as lithiumdiisopropylamide, n-butyl lithium, sodium hydride, triethylamine, andHunig's base. The reaction generally is carried out at a temperature ofabout −78° C. to about 100° C., and normally is complete within about 2hours to about 4 days. The product can be isolated by removing thesolvent, for example by evaporation under reduced pressure, and furtherpurified, if desired, by standard methods such as chromatography,crystallization, or distillation.

[0353] The 2-(phenylamino)-benzoic acid (e.g., Formula I, where R₇ ishydrogen) can be reacted with an organic or inorganic base such aspyridine, triethylamine, calcium carbonate, or sodium hydroxide toproduce a pharmaceutically acceptable salt. The free acids can also bereacted with an alcohol of the formula HOR₇ (where R₇ is other thanhydrogen, for example methyl) to produce the corresponding ester.Reaction of the benzoic acid with an alcohol can be carried out in thepresence of a coupling agent. Typical coupling reagents include2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline (EEDQ),1,3-dicyclohexylcarbodiimide (DCC), bromo-tris(pyrrolidino)-phosphoniumhexafluorophosphate (PyBrOP), and (benzotriazolyloxy) tripyrrolidinophosphonium hexafluorophosphate (PyBOP). The phenylamino benzoic acidand alcohol derivative normally are mixed in approximately equimolarquantities in an unreactive organic solvent such as dichloromethane,tetrahydrofuran, chloroform, or xylene, and an equimolar quantity of thecoupling reagent is added. A base such as triethylamine ordiisopropylethylamine can be added to act as an acid scavenger ifdesired. The coupling reaction generally is complete after about 10minutes to 2 hours, and the product is readily isolated by removing thereaction solvent, for instance by evaporation under reduced pressure,and purifying the product by standard methods such as chromatography orcrystallizations from solvents such as acetone, diethyl ether, orethanol.

[0354] The benzamides of the invention, Formula I where Z is CONR₆R₇,are readily prepared by reacting the foregoing benzoic acids with anamine of the formula HNR₆R₇. The reaction is carried out by reactingapproximately equimolar quantities of the benzoic acid and amine in anunreactive organic solvent in the presence of a coupling reagent.Typical solvents are chloroform, dichloromethane, tetrahydrofuran,benzene, toluene, and xylene. Typical coupling reagents include DCC,EEDQ, PyBrOP, and PyBOP. The reaction is generally complete after about10 minutes to about 2 hours when carried out at a temperature of about0° C. to about 60° C. The product amide is readily isolated by removingthe reaction solvent, for instance by evaporation, and furtherpurification can be accomplished by normal methods such aschromatography, crystallization, or distillation. The hydrazides(z=CONHNR₁₀R₁₁) are similarly prepared by coupling a benzoic acid with ahydrazine of the formula

H₂HNR₁₀R₁₁.

[0355] The benzyl alcohols of the invention, compounds of Formula Iwhere Z is CH₂OR₆ and R₆ is hydrogen, are readily prepared by reductionof the corresponding benzoic acid according to the following Scheme 2.

[0356] Typical reducing agents commonly employed include borane intetrahydrofuran. The reduction normally is carried out in an unreactiveorganic solvent such as tetrahydrofuran, and generally is completewithin about 2 hours to about 24 hours when conducted at a temperatureof about 0° C. to about 40° C.

[0357] The following detailed examples illustrate specific compoundsprovided by this invention.

EXAMPLE 1 4-Fluoro-2-(4-iodo-2-methylphenylamino)benzoic acid

[0358] To a stirring solution comprised of 3.16 g (0.0133 mol) of2-amino-5-iodotoluene in 5 mL of tetrahydrofuran at −78° C. was added 10mL (0.020 mol) of a 2.0 M lithium diisopropylamide intetrahydrofuran/heptane/ethenylbenzene (Aldrich) solution. The resultinggreen suspension was stirred vigorously for 15 minutes, after which timea solution of 1.00 g (0.00632 mol) of 2,4-difluorobenzoic acid in 10 mLof tetrahydrofuran was added. The reaction temperature was allowed toincrease slowly to room temperature, at which temperature it was stirredfor 2 days. The reaction mixture was concentrated. Aqueous HCl (10%) wasadded to the concentrate, and the solution was extracted withdichloromethane. The organic phase was dried (MgSO₄) and then boiledover a steambath to low volume and cooled to room temperature. Theoff-white fibers were collected by vacuum filtration, rinsed withhexanes, and vacuum-oven dried. (76° C.; ca. 10 mm of Hg) to afford 1.10g (47%) of the desired material;

[0359] mp 224-229.5° C.; ¹H NMR (400 MHz; DMSO): δ 9.72 (s, 1H), 7.97(dd, 1H, J=7.0, 8.7 Hz), 7.70 (d, 1H, J=1.5 Hz), 7.57 (dd, 1H, J=8.4,1.9 Hz), 7.17 (d, 1H, J=8.2 Hz), 6.61-6.53 (m, 2H), 2.18 (s, 3H); ¹³CNMR (100 MHz; DMSO): δ 169.87, 167.60, 165.12, 150.17, 150.05, 139.83,138.49, 136.07, 135.31, 135.20, 135.07, 125.60, 109.32, 105.09, 104.87,99.72, 99.46, 89.43, 17.52; ¹⁹F NMR (376 MHz; DMSO): δ −104.00 to−104.07 (m); IR (KBr) 1670 (C═O stretch) cm⁻¹; MS (CI) M+1=372. Analysiscalculated for C₁₄H₁₁FINO₂: C, 45.31; H, 2.99; N, 3.77. Found: C, 45.21;H, 2.77; N, 3.64.

EXAMPLES 2-30

[0360] By following the general procedure of Example 1, the followingbenzoic acids and salts of Formula (I) were prepared. Example No.Compound MP° C. 2 3,4,5-Trifluoro-2-(4-iodo-2-methyl-phenylamino)-206-210 benzoic acid 33,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic 240.5-244.5 acid 45-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 259.5-262  phenylamino)-benzoic acid 55-Chloro-2-(2-chloro-4-iodo-phenylamino)-benzoic acid 255-260 65-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 234-238 7 Sodium5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 310-320 benzoate DEC 85-Bromo-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 239.5-240   92-(2-Chloro-4-iodo-phenylamino)-5-nitro-benzoic acid 289-293 104-Fluoro-2-(3-fluoro-4-iodo-2-methyl-phenylamino)- 233-235 benzoic acid11 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-benzoic acid 264-267 122-(2-Fluoro-4-iodo-phenylamino)-5-nitro-benzoic acid 256-258 132-(4-Bromo-2-methyl-phenylamino)-4-fluoro-benzoic 218.5-220   acid 142-(2-Bromo-4-iodo-phenylamino)-5-nitro-benzoic acid 285-288 DEC 152-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro- 230-234 benzoic acid 163-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 218-221 173,4-Difluoro-2-(4-iodo-2-methoxy-phenylamino)- 230-233 benzoic acid 184-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 245-255 DEC 192-(4-Iodo-2-methyl-phenylamino)-benzoic acid 218-223 205-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 243-46  215-Iodo-2-(4-iodo-2-methyl-phenylamino)-benzoic acid 241-245 222,3,5-Trifluoro-4-(4-iodo-2-methyl-phenylamino)- 218-222 benzoic acid 234-Fluoro-2-(3-chloro-4-iodo-2-methyl-phenylamino)-   248-252.5 benzoicacid 24 2-(4-Iodo-phenylamino)-5-methoxy-benzoic acid 208-211 253-Chloro-2-(2-chloro-4-iodo-phenylamino)-benzoic acid 232-233 262-Fluoro-6-(4-iodo-2-methyl-phenylamino)-benzoic acid 179-182 274-Fluoro2-(2,3-dimethyl-4-iodo-2-methyl- 258-261 phenylamino)benzoicacid 28 5-Methyl-2-(4-iodo-2-methyl-phenylamino)-benzoic 209.5-211  acid 29 2-Chloro-6-(4-iodo-2-methyl-phenylamino)-benzoic acid 171-175 302-(4-Iodo-2-methyl-phenylamino)-4-nitro-benzoic acid 251-263

EXAMPLE 315Chloro-N-(2-hydroxyethyl)-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0361] To a stirring solution comprised of 0.1020 g (0.2632 mmol) of5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid, 0.1 mL (1.7 mmol)of ethanolamine, and 0.05 mL (0.29 mmol) of diisopropylethylamine in 5mL of a 1:1 (v/v) tetrahydrofuran-dichloromethane solution was added0.15 g (0.29 mmol) of solid PyBOP powder directly. The reaction mixturewas stirred at room temperture overnight. The solvent was removed invacuo. The crude residue was partitioned between ether (50 mL) and 10%aqueous hydrochloric acid (50 mL). The organic phase was washed with 10%aqueous sodium hydroxide (50 mL), dried SO₄) and concentrated in vacuoto afford a yellow-brown oil which was crystallized from hexanes-etherto afford 0.0831 g (73%) of a green-yellow power; mp 120-121° C.;

[0362]¹H NMR (400 MHz; CDCl₃): δ 9.11 (s, 1H), 7.56 (d, 1H, J=1.4 Hz),7.46-7.41 (m, 2H), 7.20 (dd, 1H, J=8.9, 2.4 Hz), 7.00 (t, 2H, J=9.6 Hz),6.55 (broad t, 1H), 3.86 (t, 2H, J=5.0 Hz), 3.61 (dd, 2H, J=10.1, 5.5Hz), 2.23 (s, 3H), 1.56 (broad s, 1H); IR (KBr) 3297 (O—H stretch), 1627(C═O stretch) cm⁻¹; MS (CI) M+1=431. Analysis calculated forC₁₆H₁₆ClIN₂O₂: C, 44.62; H, 3.74; N, 6.50. Found: 44.63; H, 3.67; N,6.30.

EXAMPLES 32-48

[0363] By following the general procedure of Example 31, the followingbenzamides were prepared by reacting the corresponding benzoic acid withthe corresponding amine. Example No. Compound MP° C. 324-Methoxy-N-(4-methoxy-phenyl)-3-nitro- 153.5-156   benzamide 334-Fluoro-2-(4-iodo-2-methyl-phenylamino)- 158 benzamide 344-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 102.5-104.5 methyl-benzamide35 N-Ethyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)- 90-91 benzamide 364-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N,N- oil dimethyl-benzamide 374-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(1H- 285-288tetrazol-5-yl)-benzamide DEC 38 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-180-182 benzamide 39 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N,N-137-138 dimethyl-benzamide 40 [5-Chloro-2-(4-iodo-2-methyl-phenylamino)-170-173 benzoylamino]-acetic acid 414-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 69-71 propyl-benzamide 425-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-   132-133.4phenylamino)-benzamide 43 N,N-Diethyl-4-fluoro-2-(4-iodo-2-methyl- oilphenylamino)-benzamide 444-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]- 122-124propyl}-2-(4-iodo-2-methyl-phenylamino)- benzamide 45N,N-Diethyl-2-(4-iodo-2-methyl-phenylamino)-5- 91-93 nitro-benzamide 46N-Butyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)- 97-99 benzamide 475-Chloro-N,N-diethyl-2-(4-iodo-2-methyl- 118-120 phenylamino)-benzamide48 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N,N- 142.5-144  dimethyl-benzamide

EXAMPLE 49 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzyl alcohol

[0364] 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.50 g,1.35 mmol) was dissolved in 6 mL (6 mmol) of cold 1.0 Mborane-tetrahydrofuran complex in tetrahydrofuran solution. The reactionmixture was stirred under nitrogen atmosphere at room temperatureovernight. The reaction was quenched with 80 mL of methanol.Concentration in vacuo produced a clear tan oil which was purified byMPLC. Elution with dichloromethane afforded 0.4285 g 89%) of a whitesolid; mp 99-100.5° C.;

[0365]¹H NMR (400 MHz; DMSO): δ 7.57 (d, 1H, J=1.7 Hz), 7.45 (dd, 1H,J=8.4, 1.9 Hz), 7.39 (s, 1H), 7.29 (t, 1H, J=7.5 Hz), 6.89 (d, 1H, J=8.4Hz), 6.67-6.60 (m, 1H), 5.47 (t,1H, J=5.5 Hz), 4.49 (d, 2H, 5.1 Hz),2.14 (s, 3H); IR (KBr) 3372 (O—H stretch) cm⁻¹; MS (CI) M+1=358.Analysis calculated for C₁₄H₁₃FINO: C, 47.08; H, 3.67; N, 3.92. Found:C, 47.17; H, 3.75; N, 3.72.

EXAMPLE 50-52

[0366] The following benzyl alcohols were prepared by the generalprocedure of Example 49. Example No. Compound MP° C. 50[5-Chloro-2-(4-iodo-2-methyl-phenylamino)- 82-85 phenyl]-methanol 51[2-(4-Iodo-2-methyl-phenylamino)-5-nitro-phenyl]- 126.5-128.5 methanol52 [5-Bromo-2-(4-iodo-2-methyl-phenylamino)- 60.5-63.5 phenyl]-methanol

[0367] Several invention compounds of Formula I were prepared utilizingcombinatorial synthetic techniques. The general procedure is as follows:

[0368] To a 0.8-mL autosampler vial in a metal block was added 40 μL ofa 0.5 M solution of the acid in DMF and 40 μL of the reagent amine (2 Msolution in Hunig's base and 1 M in amine in DMF). A 0.5 M solution ofPyBrop was freshly prepared and 50 μL were added to the autosamplervial. The reaction was allowed to stand for 24 hours.

[0369] The reaction mixture was transferred to a 2-dram vial and dilutedwith 2 mL of ethyl acetate. The organic layer was washed with 3 mL ofdistilled water and the water layer washed again with 2 mL of ethylacetate. The combined organic layers were allowed to evaporate todryness in an open fume hood.

[0370] The residue was taken up in 2 mL of 50% acetonitrile in water andinjected on a semi-prep reversed phase column (10 mm×25 cm, 5 μMspherical silica, pore size 115 A derivatized with C-18, the sample waseluted at 4.7 mL/min with a linear ramp to 100% acetonitrile over 8.5minutes. Elution with 100% acetonitrile continued for 8 minutes).Fractions were collected by monitoring at 214 nM. The residue wasdissolved in chloroform and transferred to a preweighed vial,evaporated, and weighed again to determine the yield.

EXAMPLES 53-206

[0371] The following compounds of Formula I were prepared bycombinatorial methodology: Example MS No. Compound M − H 535-Bromo-3,4-difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl- 510phenylamino)-benzamide 54N-(2,3-Dihydroxy-propyl)-3,4-difluoro-2-(4-iodo-2-methyl- 462phenylamino)-benzamide 555-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2- 577piperidin-1-yl-ethyl)-benzamide 563,4-Difluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl- 432phenylamino)-benzamide 57N-(2,3-Dihydroxy-propyl)-4-fluoro-2-(4-iodo-2-methyl- 444phenylamino)-benzamide 583,4-Difluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl- 446phenylamino)-benzamide 595-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N- 564(2-pyrrolidin-1-yl-ethyl)-benzamide 605-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N- 571(2-pyridin-4-yl-ethyl)-benzamide 614-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 414benzamide 62 5-Bromo-N-(3-dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-551 2-methyl-phenylamino)-benzamide 635-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N- 580(2-morpholin-4-yl-ethyl)-benzamide 643,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin- 5014-yl-ethyl)-benzamide 653,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin- 4851-yl-ethyl)-benzamide 663,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-4-yl- 493ethyl)-benzamide 67N-(3-Dimethylamino-propyl)-3,4-difluoro-2-(4-iodo-2-methyl- 473phenylamino)-benzamide 68N-Benzyl-4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide 460 692-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-hydroxy- 384ethyl)-benzamide 704-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl- 483ethyl)-benzamide 714-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl- 495propyl)-benzamide 723,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin- 5131-yl-propyl)-benzamide 734-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-thiophen-2-yl- 480ethyl)-benzamide 744-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl- 467ethyl)-benzamide 752-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-morpholin- 4534-yl-ethyl)-benzamide 765-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-N- 557pyridin-4-ylmethyl-benzamide 773,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin- 4794-ylmethyl-benzamide 782-(4-Bromo-2-methyl-phenylamino)-N-(3-dimethylamino-propyl)- 4253,4-difluoro-benzamide 794-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl- 461benzamide 80 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyridin-4-yl-475 ethyl)-benzamide 812-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-pyridin- 4454-yl-ethyl)-benzamide 822-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(3-hydroxy- 400propyl)-benzamide 832-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-pyrrolidin- 4371-yl-ethyl)-benzamide 844-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-phenethyl- 474 benzamide 852-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-thiophen- 4502-yl-ethyl)-benzamide 862-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-pyridin- 4314-ylmethyl-benzamide 872-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-phenethyl- 444 benzamide88 2-(4-Bromo-2-methyl-phenylamino)-3,4-difluoro-N-(2-piperidin- 4511-yl-ethyl)-benzamide 895-Chloro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}- 557*2-(4-iodo-2-methyl-phenylamino)- benzamide 905-Fluoro-N-{3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}- 541*2-(4-iodo-2-methyl- phenylamino)- benzamide 912-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-pyridin-4-yl methyl- 487benzamide 92 5-Bromo-N-{ 3-[4-(2-hydroxy-ethyl)-piperazin-1-yl]-propyl}-601* 2-(4-iodo-2-methyl- phenylamino)- benzamide 935-Chloro-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl- 486* phenylamino)-benzamide 945-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl- 497*ethyl)-benzamide 95(3-Hydroxy-pyrrolidin-1-yl)-[2-(4-iodo-2-methyl-phenylamino)- 4665-nitro-phenyl]-methanone 965-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl- 484*ethyl)-benzamide 97 5-Bromo-N-(2-diethylamino-ethyl)-2-(4-iodo-2-methyl-530* phenylamino)- benzamide 98N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-chloro-2-(4-iodo- 518*2-methyl- phenylamino)- benzamide 99N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-bromo-2-(4-iodo- 562*2-methyl- phenylamino)- benzamide 100[5-Bromo-2-(4-iodo-2-methyl-phenylamino)-phenyl]-(3-hydroxy- 499pyrrolidin-1-yl)-methanone 1012-(4-Iodo-2-methyl-phenylamino)-5-nitro-benzoic acid phenethyl 501 ester102 N-{3-[4-(2-Hydroxy-ethyl)-piperazin-1-yl]-propyl }-2-(4-iodo- 568*2-methyl-phenylamino)- benzamide 103[5-Chloro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-(3-hydroxy- 455pyrrolidin-1-yl)-methanone 1045-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-pyridin-4-ylmethyl- 460benzamide 1055-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl- 528*ethyl)-benzamide 1065-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl- 542*ethyl)-benzamide 1075-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-pyrrolidin-1-yl- 468*ethyl)-benzamide 1085-Chloro-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl- 472*phenylamino)-benzamide 109N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-5-fluoro-2-(4-iodo- 502*2-methyl- phenylamino)- benzamide 1105-Chloro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl- 445*phenylamino)-benzamide 1115-Chloro-N-(3-diethylamino-2-hydroxy-propyl)-2-(4-iodo- 516*2-methyl-phenylamino)- benzamide 1125-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperidin-1-yl- 482*ethyl)-benzamide 113 5-Bromo-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl-489* phenylamino)-benzamide 1145-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl- 556*propyl)-benzamide 115N-{2-[Bis-(2-hydroxy-ethyl)-amino]-ethyl}-2-(4-iodo-2-methyl- 529*phenylamino)-5-nitro-benzamide 1165-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl- 500*ethyl)-benzamide 1175-Chloro-N-(3-diethylamino-propyl)-2-(4-iodo-2-methyl- 500*phenylamino)-benzamide 1185-Chloro-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl- 514*phenylamino)-benzamide 1195-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl- 512*propyl)-benzamide 1202-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(2-piperidin-1-yl- 509*ethyl)-benzamide 1215-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-piperazin-1-yl- 544*ethyl)-benzamide 122N-(2-Diethylamino-ethyl)-5-fluoro-2-(4-iodo-2-methyl- 470*phenylamino)-benzamide 1235-Bromo-N-(3-dimethylamino-propyl)-2-(4-iodo-2-methyl- 516*phenylamino)-benzamide 124N-(3-Hydroxy-propyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 456*benzamide 125 5-Fluoro-N-(3-hydroxy-propyl)-2-(4-iodo-2-methyl- 429*phenylamino)-benzamide 126N-(3-Diethylamino-propyl)-5-fluoro-2-(4-iodo-2-methyl- 484*phenylamino)-benzamide 127N-(3-Diethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)- 511*5-nitro-benzamide 1285-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl- 544*ethyl)-benzamide 1292-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(3-piperidin-1-yl- 523*propyl)-benzamide 130[5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-(3-hydroxy- 439pyrrolidin-1-yl)-methanone 1315-Bromo-N-(2-diisopropylamino-ethyl)-2-(4-iodo-2-methyl- 558*phenylamino)-benzamide 1325-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(2-morpholin-4-yl- 484*ethyl)-benzamide 1335-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-piperidin-1-yl- 496*propyl)-benzamide 134 [5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-phenyl]-482 [4-(2-hydroxy-ethyl)-piperazin-1-yl]-methanone 135N-(3-Diethylamino-2-hydroxy-propyl)-5-fluoro-2-(4-iodo- 500*2-methyl-phenylamino)-benzamide 136[5-Chloro-2-(4-iodo-2-methyl-phenylamino)-benzoylamino]- 443 acetic acid137 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(2-pyrrolidin-1-yl- 495*ethyl)-benzamide 138N-(3-Dimethylamino-propyl)-2-(4-iodo-2-methyl-phenylamino)- 483*5-nitro-benzamide 139N-(2-Diisopropylamino-ethyl)-5-fluoro-2-(4-iodo-2-methyl- 498*phenylamino)- benzamide 1405-Fluoro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic acid S- 490phenethyl ester 141 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoicacid S- 506 phenethyl ester 1425-Bromo-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic acid S- 536 benzylester 143 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-thiobenzoic acid S-503 benzyl ester 1445-Fluoro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic acid S- 476 benzylester 145 5-Chloro-2-(4-iodo-2-methyl-phenylamino)-thiobenzoic acid S-492 benzyl ester 146N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)- 409 benzamide147 5-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 429benzamide 1485-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 413benzamide 149 N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)- 475benzamide 150 N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)- 593*benzamide 151 2-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(4-sulfamoyl-567 benzyl)-benzamide 1525-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 473benzamide 153N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-phenylamino)- 521benzamide 154N-(2-Hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 440benzamide 155 2-(4-Iodo-2-methyl-phenylamino)-N-methyl-5-nitro-N-phenyl-486 benzamide 1565-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)- 425 benzamide157 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl- 459benzamide 158 N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide409 159 N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)- 583benzamide 160 5-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-538 benzyl)-benzamide 161N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzamide 425 162N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 436 benzamide 1635-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)- 469 benzamide 1645-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl- 475benzamide 1655-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-benzyl)- 646benzamide 166 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl-598 benzyl)-benzamide 167N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide 436 1682-(4-Iodo-2-methyl-phenylamino)-5-nitro-N-(4-sulfamoyl- 565benzyl)-benzamide 169N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide 469 1705-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)- 473benzamide 171 N-Cyclopropyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)- 517benzamide 172 5-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-519 benzamide 173 N-Benzyloxy-2-(4-iodo-2-methyl-phenylamino)-5-nitro-502 benzamide 174 N-Cyclohexyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-559 benzamide 175N-Allyl-5-iodo-2-(4-iodo-2-methyl-phenylamino)-benzamide 517 1765-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)- 581benzamide 1772-(4-Iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)-5-nitro- 500benzamide 178 5-Iodo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl-567 benzamide 179 N-Cyclohexyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-451 benzamide 180 5-Chloro-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-467 benzamide 1815-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)- 533benzamide 182 5-Bromo-N-cyclohexyl-2-(4-iodo-2-methyl-phenylamino)- 511benzamide 1835-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-(3-methyl-benzyl)- 489benzamide 184 N-Cyclohexyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 478benzamide 185 N-Benzyloxy-5-bromo-2-(4-iodo-2-methyl-phenylamino)- 538benzamine 186 N-Benzyloxy-5-fluoro-2-(4-iodo-2-methyl-phenylamino)- 477benzamide 1875-Chloro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 431benzamide 1885-Bromo-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 475benzamide 189 2-(4-Iodo-2-methyl-phenylamino)-N-methyl-5-nitro-N-phenyl-488 benzamide 1905-Chloro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl- 477benzamide 191N-(2-Hydroxy-ethyl)-5-iodo-2-(4-iodo-2-methyl-phenylamino)- 523benzamide 192 5-Chloro-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-425 benzamide 193N-Allyl-5-chloro-2-(4-iodo-2-methyl-phenylamino)-benzamide 427 1945-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl- 461benzamide 195N-(2-Hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 442benzamide 1965-Fluoro-N-(2-hydroxy-ethyl)-2-(4-iodo-2-methyl-phenylamino)- 415benzamide 197 5-Bromo-N-cyclopropyl-2-(4-iodo-2-methyl-phenylamino)- 472benzamide 198 N-Cyclopropyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-411 benzamide 1995-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl- 540benzyl)-benzamide 200N-Cyclopropyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro- 438 benzamide 201N-Allyl-5-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzamide 411 202N-Benzyloxy-5-iodo-2-(4-iodo-2-methyl-phenylamino)- 585 benzamide 203N-Allyl-5-bromo-2-(4-iodo-2-methyl-phenylamino)-benzamide 472 2045-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-(4-sulfamoyl- 601benzyl)-benzamide 2055-Bromo-2-(4-iodo-2-methyl-phenylamino)-N-methyl-N-phenyl- 522 benzamide206 N-Allyl-2-(4-iodo-2-methyl-phenylamino)-5-nitro-benzamide 438

EXAMPLE 207 Preparation of[4-Chloro-2-(1H-tetrazol-5-yl)-(4-iodo-2-methyl-phenyl)-amine Step a:Preparation of 5-chloro-2-fluoro-benzaldehyde

[0372] To a solution of 1-chloro-4-fluorobenzne (13.06 g, 0.1 mol) inTHF (180 mL), at −78° C., LDA (2M solution in THF, 50 mL, 0.1 mol) wasadded drop wise. After stirring at −78° C. for 1.5 hours, DMF (8 mL) wasadded to the reaction mixture and allowed to warm up to room temperatureovernight. The reaction mixture was partitioned between water and Et₂O.The Et₂O layer was dried (MgSO₄) and the solvent removed in vacuum togive 14.95 g (94%) yield of crude dehyde: ¹H NMR (CDCl₃): δ 10.3 (s,—C(═O)H).

Step b: Preparation of 5-chloro-2-fluoro-benzaldehyde oxime

[0373] A solution of 5-chloro-2-fluoro-benzaldehyde (10 g, 0.0631 mol),hydroxylamine hydrochloride (6.57 g, 0.0946 mol) and pyridine (8.3 mL,0.1010 mol) in EtOH (100 mL) was heated at 75° C. (oil bath temperature)for 1 hour and the solvent removed under vacuum to give an oil. The oilwas partitioned between water and CH₂Cl₂. The CH₂Cl₂ layer was dried(MgSO₄) and the solvent removed under vacuum to give crude aldoxime as asolid. The solid was purified by medium pressure liquid chromatographyon silica. Elution with CH₂Cl₂ gave 4.87 g (28%) of the aldoxime aswhite solid: mp 95-97° C.;

[0374] Analysis calculated for C₇H₅NOFCl: C, 48.44; H, 2.90; N, 8.07.Found: C, 48.55; H, 2.69, N, 7.90.

Step c: Preparation of 5-chloro-2-fluoro-benzonirile

[0375] A solution of the 5-chloro-2-fluoro-benzaldehyde oxime (3.15 g,0.0182 mol) in acetic anhydride (150 mL) was refluxed for 16 hours. Thereaction mixture was cooled to room temperature and poured intosaturated aqueous NaHCO₃ (200 mL) solution. The mixture was extractedwith Et₂O. The Et₂O layer was dried (K₂CO₃) and the solvent removed togive the product as an oily solid. The product was used without furtherpurification in the next step.

Step d: Preparation of 5-(5-chloro-2-fluoro-phenyl)-1H-tetrazole

[0376] A mixture of 5-chloro-2-fluoro-benzonitrile (2.84 g, 0.01823mol), butanol (15 mL), sodium azide (1.543 g, 0.0237 mol), acetic acid(1.36 mL, 0.0237 mol) was refluxed for 24 hours. The reaction mixturewas cooled to room temperature, additional 1.543 g sodium azide added,and the reaction mixture refluxed for additional 24 hours. After coolingto room temperature, Et₂O (100 mL) and 10% aqueous NaOH (200 mL) wereadded sequentially. The mixture was vigorously stirred. The aqueouslayer was separated, cooled with ice-methanol bath (−15° C.) andacidified to pH 1 with conc. HCl. A gray solid precipitated. The solidwas dried in vacuum at 50° C. to give 1.76 g (49%) of5-(5-chloro-2-fluoro-phenyl)-1H-tetrazole: mp partial melt at 110° C.,complete melting at 124° C.);

[0377]¹H (400 Mz, CDCl₃): δ 8.19-8.08 (m, 1H), 7.77-7.71 (m, 1H),7.61-7.52 (m, 1H); ¹³C (100 Mz, CDCl₃): δ 159.00, 156.49, 140.88,133.02, 132.93, 130.73, 129.23, 129.21, 129.08, 126.05, 118.96, 118.73,114.50; MS (CI) M+1=199 (100), M=198 (6).

Step e: Preparation of[4-Chloro-2-(1H-tetrazol-5-yl)-(4-iodo-2-methyl-phenyl)-amine

[0378] To a solution of 2-methyl-4-iodoaniline (3.52 g, 0.0151 mol) inTHF (25 mL) at −78° C., LDA (2 molar solution in THF, 11.33 mL, 0.02267mol) was added dropwise. After stirring for 0.5 hours, a solution of1-(tetrazol-5-yl)-2-fluoro-5-chlorobenzene (1.5 g, 0.00756 mol) in THF(15 mL) was added dropwise. The reaction was stirred for 16 hours as itwarmed up to room temperature. The reaction mixture was quenched withaqueous conc. NH₄Cl solution and extracted with CH₂Cl₂. The organiclayer was dried (MgSO₄) and the solvent removed giving a crude productas an oil. The oil with CH₂Cl₂→CH₂Cl₂:MeOH (9.7:0.3) gave 1.5 g (48%) ofthe desired product:

[0379] mp 205-208° C.; ¹H (400 Mz, DMSO): δ 9.13 (s, 1H), 8.00-7.99 (s,1H), 7.69 (s, 1H), 7.55-7.52 (m, 1H), 7.43-7.40 (m, 1H), 7.12-7.05 (m,1H), 2.24 (s, 3H); ¹³C (100 Mz, CDCl₃): δ 141.87, 139.28, 138.88,135.47, 133.71, 131.65, 128.15, 123.69, 121.94, 116.68, 87.79, 17.22; MS(CI) M+2=413 (44), M+1=412 (85), M=411 (100). Analysis calculated forC₁₄H₁₁N₅ClI.0.5H₂O: C, 39.97; H, 2.87; N, 16.65. Found: C, 38.87, H,2.77; N, 16.47.

[0380] The following tetrazole substituted phenylamines were prepared byfollowing the general procedure of Example 207.

EXAMPLE 208 (4-iodo-2-methyl-phenyl)-[2-(1H-tetrazol-5-yl)-phenyl]amine,mp 231° C. (dec) EXAMPLE 209[4-nitro-2-(1H-tetrazol-5-yl)-(4-iodo-2-methyl-phenyl)-amine, mp205-208° C.

[0381] The 4-bromo and 4-iodo phenylamino benzhydroxamic acidderivatives of Formula II can be prepared from commercially availablestarting materials utilizing synthetic methodologies well-known to thoseskilled in organic chemistry. A typical synthesis is carried out byreacting a 4-bromo or 4-iodo aniline with a benzoic acid having aleaving group at the 2-position to give a phenylamino benzoic acid, andthen reacting the benzoic acid phenylamino derivative with ahydroxylamine derivative (Scheme 3), where L is a leaving group, forexample halo such as fluoro, chloro, bromo or iodo, or an activatedhydroxy group such as a diethylphosphate, trimethylsilyloxy,p-nitrophenoxy, or phenylsulfonoxy.

[0382] The reaction of aniline and the benzoic acid derivative generallyis accomplished by mixing the benzoic acid with an equimolar quantity orexcess of the aniline in an unreactive organic solvent such astetrahydrofuran, or toluene, in the presence of a base such as lithiumdiisopropylamide, n-butyl lithium, sodium hydride, and sodium amide. Thereaction generally is carried out at a temperature of about −78° C. toabout 25° C., and normally is complete within about 2 hours to about 4days. The product can be isolated by removing the solvent, for exampleby evaporation under reduced pressure, and further purified, if desired,by standard methods such as chromatography, crystallization, ordistillation.

[0383] The phenylamino benzoic acid next is reacted with a hydroxylaminederivative HNR_(6a)OR_(7a) in the presence of a peptide couplingreagent. Hydroxylamine derivatives that can be employed includemethoxylamine, N-ethyl-isopropoxy amine, and tetrahydro-oxazine. Typicalcoupling reagents include 2-ethoxy-1-ethoxycarbonyl-1,2-dihydroquinoline(EEDQ), 1,3-dicyclohexylcarbodiimide (DCC),bromo-tris(pyrrolidino)-phosphonium hexafluorophosphate (PyBrOP) and(benzotriazolyloxy)tripyrrolidino phosphonium hexafluorophosphate(PyBOP). The phenylamino benzoic acid and hydroxylamino derivativenormally are mixed in approximately equimolar quantities in anunreactive organic solvent such as dichloromethane, tetrahydrofuran,chloroform, or xylene, and an equimolar quantity of the coupling reagentis added. A base such as triethylamine or diisopropylethylamine can beadded to act as an acid scavenger if desired. The coupling reactiongenerally is complete after about 10 minutes to 2 hours, and the productis readily isolated by removing the reaction solvent, for instance byevaporation under reduced pressure, and purifying the product bystandard methods such as chromatography or crystallizations fromsolvents such as acetone, diethyl ether, or ethanol. An alternativemethod for making the invention compounds involves first converting abenzoic acid to a hydroxamic acid derivative, and then reacting thehydroxamic acid derivative with an aniline. This synthetic sequence isdepicted in Scheme 4, where L is a leaving group. The general reactionconditions for both of the steps in Scheme 4 are the same as thosedescribed above for Scheme 3.

[0384] Yet another method for making invention compounds comprisesreacting a phenylamino benzhydroxamic acid with an ester forming groupas depicted in Scheme 5, where L is a leaving group such as halo, and abase is triethylamine or diisopropylamine.

[0385] The synthesis of compounds of Formula (II) is further illustratedby the following detailed examples.

EXAMPLE 1a 4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide(a) Preparation of 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0386] To a stirred solution containing 3.16 g (0.0133 mol) of2-amino-5-iodotoluene in 5 mL of tetrahydrofuran at −78° C. was added 10mL (0.020 mol) of a 2.0 M lithium diisopropylamide intetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resultinggreen suspension was stirred vigorously for 15 minutes, after which timea solution of 1.00 g (0.00632 mol) of 2,4-difluorobenzoic acid in 10 mLof tetrahydrofuran was added. The reaction temperature was allowed toincrease slowly to room temperature, at which temperature the mixturewas stirred for 2 days. The reaction mixture was concentrated byevaporation of the solvent under reduced pressure. Aqueous HCl (10%) wasadded to the concentrate, and the solution was extracted withdichloromethane. The organic phase was dried (MgSO₄) and thenconcentrated over a steambath to low volume (10 mL) and cooled to roomtemperature. The off-white fibers which formed were collected by vacuumfiltration, rinsed with hexane, and dried in a vacuum-oven (76° C.; ca.10 mm of Hg) to afford 1.10 g (47%) of the desired material; mp224-229.5° C.;

[0387]¹H NMR (400 MHz, DMSO): δ 9.72 (s, 1H), 7.97 (dd, 1H, J=7.0, 8.7Hz), 7.70 (d, 1H, J=1.5 Hz), 7.57 (dd, 1H, J=8.4, 1.9 Hz), 7.17 (d, 1H,J=8.2 Hz), 6.61-6.53 (m, 2H), 2.18 (s, 3H); ¹³C NMR (100 MHz, DMSO): δ169.87, 166.36 (d, J_(C—F)=249.4 Hz), 150.11 (d, J_(C—F)=11.4 Hz),139.83, 138.49, 136.07, 135.26 (d, J_(C—F)=1.5 Hz), 135.07, 125.60,109.32, 104.98 (d, J_(C—F)=21.1 Hz), 99.54 (d, J_(C—F)=26.0 Hz), 89.43,17.52; ¹⁹F NMR (376 MHz, DMSO): δ −104.00 to −104.07 (m); IR (KBr) 1670(C═O stretch)cm⁻¹; MS (CI) M+1=372. Analysis calculated for C₁₄H₁₁FINO₂:C, 45.31; H, 2.99; N, 3.77. Found: C, 45.21; H, 2.77; N, 3.64.

(b) Preparation of4-Fluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0388] To a stirred solution of4-fluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.6495 g,0.001750 mol), O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.2590 g,0.002211 mol), and diisopropylethylamine (0.40 mL, 0.0023 mol) in 31 mLof an equivolume tetrahydrofuran-dichloromethane solution was added 1.18g (0.00227 mol) of solid PyBOP ([benzotriazolyloxy]tripyrrolidinophosphonium hexafluorophosphate, Advanced ChemTech) directly. Thereaction mixture was stirred for 30 minutes after which time it wasconcentrated in vacuo. The brown oil was treated with 10% aqueoushydrochloric acid. The suspension was extracted with ether. The organicextraction was washed with 10% sodium hydroxide followed by another 10%hydrochloric acid wash, was dried (MgSO₄) and concentrated in vacuo toafford 1.0 g of a light-brown foam. This intermediate was dissolved in25 mL of ethanolic hydrogen chloride, and the solution was allowed tostand at room temperature for 15 minutes. The reaction mixture wasconcentrated in vacuo to a brown oil that was purified by flash silicachromatography. Elution with a gradient (100% dichloromethane to 0.6%methanol in dichloromethane) afforded 0.2284 g of a light-brown viscousoil. Scratching with pentane-hexanes and drying under high vacuumafforded 0.1541 g (23%) of an off-white foam; mp 61-75° C.;

[0389]¹H NMR (400 MHz, DMSO): δ 11.34 (s, 1H), 9.68 (s, 1H), 9.18 (s,1H), 7.65 (d, 1H, J=1.5 Hz), 7.58 (dd, 1H, J=8.7, 6.8 Hz), 7.52 (dd, 1H,J=8.4, 1.9 Hz), 7.15 (d, 1H, J=8.4 Hz), 6.74 (dd, 1H, J=11.8, 2.4 Hz),6.62 (ddd, 1H, J=8.4, 8.4, 2.7 Hz), 2.18 (s, 3H); ¹³C NMR (100 MHz,DMSO): δ 165.91, 164.36 (d, J_(C—F)=247.1 Hz), 146.78, 139.18, 138.77,135.43, 132.64, 130.60 (d, J_(C—F)=11.5 Hz), 122.23, 112.52, 104.72 (d,J=22.1 Hz), 100.45 (d, J_(C—F)=25.2 Hz), 86.77, 17.03; ¹⁹F NMR (376 MHz,DMSO): δ −107.20 to −107.27 (m); IR (KBr) 3307 (broad, O—H stretch),1636 (C═O stretch) cm⁻¹; MS (CI) M+1=387. Analysis calculated forC₁₄H₁₂FIN₂O₂: C, 43.54; H, 3.13; N, 7.25. Found: C, 43.62; H, 3.24; N,6.98.

EXAMPLE 2a5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide(a) Preparation of 5-Bromo-2,3,4-trifluorobenzoic acid

[0390] To a stirred solution comprised of 1-bromo-2,3,4-trifluorobenzene(Aldrich, 99%; 5.30 g, 0.0249 mol) in 95 mL of anhydrous tetrahydrofurancooled to −78° C. was slowly added 12.5 mL of 2.0 M lithiumdiisopropylamide in heptane/tetrahydrofuran/ethylbenzene solution(Aldrich). The mixture was stirred for 1 hour and transferred by canulainto 700 mL of a stirred saturated ethereal carbon dioxide solutioncooled to −78° C. The cold bath was removed, and the reaction mixturewas stirred for 18 hours at ambient temperature. Dilute (10%) aqueoushydrochloric acid (ca. 500 mL) was poured into the reaction mixture, andthe mixture was subsequently concentrated on a rotary evaporator to acrude solid. The solid product was partitioned between diethyl ether(150 mL) and aq. HCl (330 mL, pH 0). The aqueous phase was extractedwith a second portion (100 mL) of diethyl ether, and the combinedethereal extracts were washed with 5% aqueous sodium hydroxide (200 mL)and water (100 mL, pH 12). These combined alkaline aqueous extractionswere acidified to pH 0 with concentrated aqueous hydrochloric acid. Theresulting suspension was extracted with ether (2×200 mL). The combinedorganic extracts were dried (MgSO₄), concentrated in vacuo, andsubjected to high vacuum until constant mass was achieved to afford 5.60g (88% yield) of an off-white powder; mp 139-142.5° C.;

[0391]¹H NMR (400 MHz, DMSO): δ 13.97 (broad s, 1H, 8.00-7.96 (m, 1H);¹³C NMR (100 MHz, DMSO): δ 162.96, 129.34, 118.47, 104.54 (d,J_(C—F)=22.9 Hz); ¹⁹F NMR (376 MHz, DMSO): δ −120.20 to −120.31 (m),−131.75 to −131.86 (m), −154.95 to −155.07 (m); IR (KBr) 1696 (C═Ostretch)cm⁻¹; MS (CI) M+1=255. Analysis calculated for C₇₄H₂₁BrF₃O₂: C,32.97; H, 0.79; N, 0.00; Br, 31.34; F, 22.35. Found: C, 33.18; H, 0.64;N, 0.01; Br, 30.14; F, 22.75.

(b) Preparation of5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid

[0392] To a stirred solution comprised of 1.88 g (0.00791 mol) of2-amino-5-iodotoluene in 10 mL of tetrahydrofuran at −78° C. was added 6mL (0.012 mol) of a 2.0 M lithium diisopropylamide intetrahydrofuran/heptane/ethylbenzene (Aldrich) solution. The resultinggreen suspension was stirred vigorously for 10 minutes, after which timea solution of 1.00 g (0.00392 mol) of 5-bromo-2,3,4-trifluorobenzoicacid in 15 mL of tetrahydrofuran was added. The cold bath wassubsequently removed, and the reaction mixture stirred for 18 hours. Themixture was concentrated, and the concentrate was treated with 100 mL ofdilute (10%) aqueous hydrochloric acid. The resulting suspension wasextracted with ether (2×150 mL), and the combined organic extractionswere dried (MgSO₄) and concentrated in vacuo to give an orange solid.The solid was triturated with boiling dichloromethane, cooled to ambienttemperature, and collected by filtration. The solid was rinsed withdichloromethane, and dried in the vacuum-oven (80° C.) to afford 1.39 g(76%) of a yellow-green powder; mp 259.5-262° C.;

[0393]¹H NMR (400 MHz, DMSO): δ 9.03 (s, 1H), 7.99 (dd, 1H, J=7.5, 1.9Hz), 7.57 (dd, 1H, J=1.5 Hz), 7.42 (dd, 1H, J=8.4, 1.9 Hz), 6.70 (dd,1H, J=8.4, 6.0 Hz), 2.24 (s, 3H); ¹⁹F NMR (376 MHz, DMSO): δ −123.40 to−123.47 (m); −139.00 to −139.14 (m); IR (KBr) 1667 (C═O stretch)cm⁻¹; MS(CI) M+1=469. Analysis calculated for C₁₄H₉BrF₂INO₂: C, 35.93; H, 1.94;N, 2.99; Br, 17.07; F, 8.12; I, 27.11. Found: C, 36.15; H, 1.91; N,2.70; Br, 16.40; F, 8.46; I, 26.05.

(c) Preparation of5-Bromo-3,4-difluoro-N-hydroxy-2-(4-iodo-2-methyl-phenylamino)-benzamide

[0394] To a stirred solution comprised of5-bromo-3,4-difluoro-2-(4-iodo-2-methyl-phenylamino)-benzoic acid (0.51g, 0.0011 mol), O-(tetrahydro-2H-pyran-2-yl)-hydroxylamine (0.15 g,0.0013 mol), and diisopropylethylamine (0.25 mL, 0.0014 mol) in 20 mL ofan equivolume tetrahydrofuran-dichloromethane solution was added 0.6794g (0.001306 mol) of solid PyBOP (Advanced ChemTech) directly. Thereaction mixture was stirred at 24° C. for 10 minutes, and then wasconcentrated to dryness in vacuo. The concentrate was suspended in 100mL of 10% aqueous hydrochloric acid. The suspension was extracted with125 mL of diethyl ether. The ether layer was separated, washed with 75mL of 10% aqueous sodium hydroxide, and then with 100 mL of dilute acid.The ether solution was dried (MgSO₄) and concentrated in vacuo to afford0.62 g (100%) of an off-white foam. The foam was dissolved in ca. 15 mLof methanolic hydrogen chloride. After 5 minutes, the solution wasconcentrated in vacuo to an oil, and the oil was purified by flashsilica chromatography. Elution with dichloromethane:dichloromethane-methanol (99:1) afforded 0.2233 g (42%) of a yellowpowder. The powder was dissolved in diethyl ether and washed with dilutehydrochloric acid. The organic phase was dried (MgSO₄) and concentratedin vacuo to afford 0.200 g of a foam. This product was triturated withpentane to afford 0.1525 g of a powder that was repurified by flashsilica chromatography. Elution with dichloromethane afforded 0.0783 g(15%) of an analytically pure title compound, mp 80-90° C.;

[0395]¹H NMR (400 MHz, DMSO): δ 11.53 (s, 1H), 9.38 (s, 1H), 8.82 (s,1H), 7.70 (dd, 1H, J=7.0, 1.9 Hz), 7.53 (s, 1H), 7.37 (dd, 1H, J=8.4,1.9 Hz), 6.55 (dd, 1H, J=8.2, 6.5 Hz), 2.22 (s, 3H); ¹⁹F NMR (376 MHz,DMSO): δ −126.24 to −126.29 (m), −137.71 to −137.77 (m); IR (KBr) 3346(broad, O—H stretch), 1651 (C═O stretch)cm⁻¹; MS (CI) M+1=484. Analysiscalculated for C₁₄H₁₀BrF₂IN₂O₂: C, 34.81; H, 2.09; N, 5.80. Found: C,34.53; H, 1.73; N, 5.52.

[0396] Examples 3a to 12a in the table below were prepared by thegeneral procedure of Examples 1a and 2a.

EXAMPLES 13a-77a

[0397] Examples 13a to 77a were prepared utilizing combinatorialsynthetic methodology by reacting appropriately substituted phenylaminobenzoic acids (e.g., as shown in Scheme 1) and hydroxylamines (e.g.,(NHR_(6a))—O—R_(7a)). A general method is given below:

[0398] To a 0.8-mL autosampler vial in a metal block was added 40 μL ofa 0.5 M solution of the acid in DMF and 40 μL of the hydroxylamine (2 Msolution in Hunig's base and 1 M in amine in DMF). A 0.5 M solution ofPyBrOP was freshly prepared, and 50 μL were added to the autosamplervial. The reaction was allowed to stand for 24 hours.

[0399] The reaction mixture was transferred to a 2-dram vial and dilutedwith 2 mL of ethyl acetate. The organic layer was washed with 3 mL ofdistilled water and the water layer washed again with 2 mL of ethylacetate. The combined organic layers were allowed to evaporate todryness in an open fume hood.

[0400] The residue was taken up in 2 mL of 50% acetonitrile in water andinjected on a semi-prep reversed phase column (10 mm×25 cm, 5 μMspherical silica, pore Size 115 A derivatized with C-18, the sample waseluted at 4.7 mL/min with a linear ramp to 100% acetonitrile over 8.5minutes. Elution with 100% acetonitrile continued for 8 minutes.)Fractions were collected by monitoring at 214 nM. The desired fractionswere evaporated using a Zymark Turbovap. The product was dissolved inchloroform and transferred to a preweighed vial, evaporated, and weighedagain to determine the yield. The structure was confirmed by massspectroscopy.

EXAMPLES 3a-77a

[0401] Example Melting MS No. Compound Point (° C.) (M − H⁺)  3a2-(4-bromo-2-methyl-phenylamino)-4-fluoro-N- 56-75 523 hydroxy-benzamidedec  4a 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 65 phenylamino)-benzamidedec  5a 5-Chloro-N-hydroxy-2-(4-iodo-2-methyl- 62-67phenylamino)-N-methyl-benzamide  6a5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N- 105-108(terahydropyran-2-yloxy)benzamide  7a5-Chloro-2-(4-iodo-2-methyl-phenylamino)-N- 64-68 methoxybenzamide  8a4-Fluoro-N-hydroxy-2-(4-fluoro-2-methyl- 119-135 phenylamino)-benzamide 9a 4-Fluoro-N-hydroxy-2-(2-methyl phenylamino)- 101-103 benzamide 10a4-Fluoro-2-(4-fluor-2-methyl-phenylamino)-N- 142-146(terahydropyran-2-yloxy)benzamide 11a4-Fluoro-N-hydroxy-2-(4-cluoro-2-methyl- 133.5-135  phenylamino)-benzamide 12a 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N-  107-109.5 phenylmethoxy-benzamide 13a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 399 methoxy-benzamide 14a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 417 N-methoxy-benzamide15a 2-(4-Bromo-2-methyl-phenylamino)- 3693,4-difluoro-N-methoxy-benzamide 16a2-(4-Bromo-2-methyl-phenylamino)-N-ethoxy- 342* 3,4-difluoro-benzamide(M − EtO) 17a 5-Bromo-N-ethoxy-3,4-difluoro-2-(4-iodo- 5092-methyl-phenylamino)-benzamide 18a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445 N-isopropoxy-benzamide19a 2-(4-Bromo-2-methyl-phenylamino)- 3973,4-difluoro-N-isopropoxy-benzamide 20a4-Fluoro-N-(furan-3-ylmethoxy)-2-(4-iodo- 4652-methyl-phenylamino)-benzamide 21a3,4-Difluoro-N-(furan-3-ylmethoxy)-2-(4-iodo- 4832-methyl-phenyIamino)-benzamide 22a 2-(4-Bromo-2-methyl-phenylamino)-435 3,4-difluoro-N-(furan-3-ylmethoxy)-benzamide 23a5-Bromo-3,4-difluoro-N-(furan-3-ylmethoxy)- 5612-(4-iodo-2-methyl-phenylamino)-benzamide 24a5-Bromo-N-(but-2-enyloxy)-3,4-difluoro- 5362-(4-iodo-2-methyl-phenylamino)-benzamide 25a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 423(prop-2-ynyloxy)-benzamide 26a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 441N-(prop-2-ynyloxy)-benzamide 27a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 455N-(1-methyl-prop-2-ynyloxy)-benzamide 28a2-(4-Bromo-2-methyl-phenylamino)- 4073,4-difluoro-N-(1-methyl-prop-2-ynyloxy)- benzamide 29aN-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 4552-methyl-phenylamino)-benzamide 30a2-(4-Bromo-2-methyl-phenylamino)-N-(but- 4073-ynyloxy)-3,4-difluoro-benzamide 31a5-Bromo-N-(but-3-ynyloxy)-3,4-difluoro- 5332-(4-iodo-2-methyl-phenylamino)-benzamide 32a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 517N-(3-phenyl-prop-2-ynyloxy)-benzamide 33a3,4-Difluoro-2-(4-bromo-2-methyl- 469phenylamino)-N-(3-phenyl-prop-2-ynyloxy)- benzamide 34a3,4-Difluoro-N-[3-(3-fluoro-phenyl)-prop- 5352-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)- benzamide 35a2-(4-Bromo-2-methyl-phenylamino)- 4873,4-difluoro-N-[3-(3-fluoro-phenyl)-prop- 2-ynyloxy]-benzamide 36a3,4-Difluoro-N-[3-(2-fluoro-phenyl)-prop- 5352-ynyloxy]-2-(4-iodo-2-methyl-phenylamino)- benzamide 37a5-Bromo-3,4-difluoro-N-[3-(2-fluoro-phenyl)- 613prop-2-ynyloxy]-2-(4-iodo-2-methyl- phenylamino)-benzamide 38a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 557*N-(3-methyl-5-phenyl-pent-2-en-4-ynyloxy)- *(M + H) benzamide 39a2-(4-Bromo-2-methyl-phenylamino)- 5103,4-difluoro-N-(3-methyl-5-phenyl-pent-2-en- 4-ynyloxy)-benzamide 40aN-Ethoxy-3,4-difluoro-2-(4-iodo-2-methyl- 431 phenylamino)-benzamide 41a2-(4-Bromo-2-methyl-phenylamino)-N-ethoxy- 383 3,4-difluoro-benzamide42a 4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 427 propoxy-benzamide43a 3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445N-propoxy-benzamide 44a 2-(4-Bromo-2-methyl-phenylamino)- 3973,4-difluoro-N-propoxy-benzamide 45a5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523phenylamino)-N-propoxy-benzamide 46a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 427 isopropoxy-benzamide 47a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 445 N-isopropoxy-benzamide48a 2-(4-Bromo-2-methyl-phenylamino)- 3973,4-difluoro-N-isopropoxy-benzamide 49a5-Bromo-3,4-difluoro-2-(4-iodo-2-methyl- 523phenylamino)-N-isopropoxy-benzamide 50aN-Cyclobutyloxy-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 51a 2-(4-Bromo-2-methyl-phenylamino)-N-409 cyclobutyloxy-3,4-difluoro-benzamide 52aN-Cyclopentyloxy-4-fluoro-2-(4-iodo-2-methyl- 453 phenylamino)-benzamide53a N-Cyclopentyloxy-3,4-difluoro-2-(4-iodo- 4712-methyl-phenylamino)-benzamide 54a 2-(4-Bromo-2-methyl-phenylamino)-N-423 cyclopentyloxy-3,4-difluoro-benzamide 55aN-Cyclopropylmethoxy-4-fluoro-2-(4-iodo- 4392-methyl-phenylamino)-benzamide 56aN-Cyclopropylmethoxy-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 57a 2-(4-Bromo-2-methyl-phenylamino)-N-409 cyclopropylmethoxy-3,4-difluoro-benzamide 58a5-Bromo-N-cyclopropylmethoxy-3,4-difluoro- 4352-(4-iodo-2-methyl-phenylamino) 59a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 505(2-phenoxy-ethoxy)-benzamide 60a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 523N-(2-phenoxy-ethoxy)-benzamide 61a 2-(4-Bromo-2-methyl-phenylamino)- 4753,4-difluoro-N-(2-phenoxy-ethoxy)-benzamide 62a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 481(thiophen-2-ylmethoxy)-benzamide 63a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 499N-(thiophen-2-ylmethoxy)-benzamide 64a 2-(4-Bromo-2-methyl-phenylamino)-451 3,4-difluoro-N-(thiophen-2-ylmethoxy)- benzamide 65a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 439(2-methyl-allyloxy)-benzamide 66a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 457N-(2-methyl-allyloxy)-benzamide 67a 2-(4-Bromo-2-methyl-phenylamino)-410 3,4-difluoro-N-(2-methyl-allyloxy)-benzamide 68aN-(But-2-enyloxy)-4-fluoro-2-(4-iodo-2-methyl- 439phenylamino)-benzamide 69a N-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 70a2-(4-Bromo-2-methyl-phenylarnino)-N-(but- 4102-enyloxy)-3,4-difluoro-benzamide 71a3,4-Difluoro-2-(4-iodo-2-methyl-phenylamino)- 441N-(prop-2-ynyloxy)-benzamide 72aN-(But-3-ynyloxy)-3,4-difluoro-2-(4-iodo- 4552-methyl-phenylamino)-benzamide 73a 2-(4-Bromo-2-methyl-phenylamino)-N-449 (4,4-dimethyl-pent-2-ynyloxy)-3,4-difluoro- benzamide 74aN-(But-2-enyloxy)-3,4-difluoro-2-(4-iodo- 4572-methyl-phenylamino)-benzamide 75a2-(4-Bromo-2-methyl-phenylamino)-N-(but- 4102-enyloxy)-3,4-difluoro-benzamide 76aN-(3-tert-butyl-propyn-2-yl)oxy-4-fluoro- 4792-(4-iodo-2-methyl-phenylamino)-benzamide 77a4-Fluoro-2-(4-iodo-2-methyl-phenylamino)-N- 577 phenylmethoxy-benzamide

[0402] PHYSICAL DATA FOR SELECTED COMPOUNDS PD 0171984 mp 80-90° C. PD0184161 mp 174-175° C. PD 0203311 mp 141-144° C. PD 0297189 mp 167-169°C. ¹H—NMR (400 MHz; DMSO) δ 11.70 (s, 1H), 8.59 (s, 1H), 7.55 (s, 1H),7.43 (d, 1H, J=6.5 Hz), 7.27 (d, 1H, J=8.7 Hz), 6.46 (m, 1H), 3.42 (d,2H, J=7.0 Hz), 0.84 (m, 1H), 0.27 (m, 2H), 0.00 (m, 2H) PD 0297190 mp125.5-133° C. ¹H—NMR (400 MHz; DMSO) δ 11.48 (s, 1H), 8.32 (s, 1H), 7.34(d, 1H, J=7.5 Hz), 7.28 (d, 2H, J=8.2 Hz), 6.48 (d, 2H, J=7.7 Hz), 3.32(d, 2H, J=6.8 Hz), 0.81 (m, 1H), 0.28 (m, 2H), 0.00 (m, 2H) PD 0296771mp 266.7-268.9° C. ¹H—NMR (400 MHz; DMSO) δ 13.85 (broad s, 1H), 8.99(s, 1H), 7.87 (dd, 1H, J=7.9, 2.1 Hz), 7.55 (d, 2H, J=8.6 Hz), 6.82 (dd,2H, J=8.7, 2.8 Hz) PD 0296770 mp 293.2-296.3° C. ¹H—NMR (400 MHz; DMSO)δ 14.05 (broad s, 1H), 9.21 (s, 1H), 7.93 (dd, 1H, J=7.8, 2.2 Hz), 7.82(d, 1H, J=1.9 Hz), 7.54 (dd, 1H, J=8.6, 1.9 Hz), 6.82 (dd, 1H, J=8.6,6.7 Hz) PD 0296767 mp 249-251° C. ¹H—NMR (400 MHz; DMSO) δ 13.99 (broads, 1H), 9.01 (s, 1H), 7.90 (dd, 1H, J=7.9, 2.3 Hz), 7.58 (d, 1H, J=1.6Hz), 7.42 (dd, 1H, J=8.4, 1.9 Hz), 6.69 (dd, 1H, J=8.4, 6.0 Hz), 2.24(s, 3H) PD298127 mp 127-135° C. 5-chloro-N-cyclopropylmethoxy-3,4-difluoro-2-[4-iodo-2-methyl phenylamino]benzamide Proton NMR(440 MHz; DMSO) δ 11.64 (s, 1H), 8.28 (s, 1H), 7.38 (dd, 1H, J=7.6, 1.7Hz), 7.31 (d, 1H, J=1.2 Hz), 7.15 (dd, 1H, J=8.5, 1.7 Hz), 3.35 (d, 2H,J=7.3 Hz), 2.01 (s, 3H), 0.83 (m, 1H), 0.28 (m, 2H), 0.01 (m,2H)

Biological Assays

[0403] The ability of MEK inhibitors described above to prevent andtreat asthma has been demonstrated in three different assays: (1)inhibition of antigen-induced interleukin-5 (IL-5) production in vitro,(2) inhibition of the passive-transfer of eosinophilic lung inflammationin vivo, and (3) inhibition of active eosinophilic lung inflammation invivo. For each of these assays, female C57BL/6 mice obtained from theJackson Laboratory (Bar Harbor, Me.) were given an intraperitoneal(i.p.) injection of ovalbumin (OVA, Grade V, Sigma Chemical Company, St.Louis, Mo.) adsorbed to aluminum hydroxide (10 μg OVA+9 mg aluminumhydroxide in 200 μL saline). This sensitizes OVA-specific lymphocytesfor subsequent restimulation either in vivo or in vitro.

[0404] The first set of experiments was designed to determine whetherthe MEK inhibitors could prevent antigen-induced production of IL-5 bythe OVA-primed splenocytes in vitro. IL-5 is required for thedifferentiation, migration, and survival of pulmonary eosinophils, whichare thought to be responsible for much of the pathology associated withhuman asthma. In order to examine the effects of MEK inhibitors on IL-5production, OVA-sensitized mice were sacrificed by cervical dislocation14 days after sensitization (Day 14), the spleens were excised anddisaggregated, and the erythrocytes were lysed. The splenocytes werewashed and resuspended at 5×10⁶ cells/mL in complete medium consistingof RPMI 1640 (Gibco BRL, Gaithersburg, Md.) with 10% heat-inactivatedfetal calf serum (Hyclone, Logan, Utah), 55 μM 2-mercaptoethanol, 50U/mL penicillin G, 50 μg/mL streptomycin sulfate, and 2 mM L-glutamine(Gibco BRL). The splenocytes were then cultured at 37° C. in thepresence of 200 μg/mL OVA. MEK inhibitors were also added to thecultures from sterile 10 mM stock solutions (in DMSO). After 3 days, theculture medium was recovered and assayed for IL-5 by specific ELISA. Theresults of the analysis of IL-5 inhibition are presented in Table 1. AllMEK inhibitors tested were found to potently inhibit antigen-inducedIL-5 production. TABLE 1 The Effects of MEK Inhibitors on Antigen-Induced IL-5 Production MEK Inhibitor IC₅₀ (nM) PD 184386 23 PD 171984117 PD 170611 1,121 PD 184161 1,147 PD 177168 1,205 PD 184352 1,622 PD098059 17,440

[0405] When OVA-sensitized spleen cells are restimulated with OVA invitro for 3 days, as described above, the spleen cells not only produceIL-5, but also acquire the ability to induce eosinophilic lunginflammation when transferred into naïve recipient mice. The criticalcell type responsible for this adoptively-transferred activity isthought to be IL-5-producing T lymphocytes. Because the results of thefirst set of experiments indicated that the MEK inhibitors inhibitedIL-5 production by cultured splenocytes, a second set of experiments wasinitiated to determine whether the MEK inhibitor-treated cells werecapable of transferring eosinophilic lung inflammation to naïve mice.Splenocytes from OVA restimulation cultures, with or without theaddition of MEK inhibitors were harvested after 3 days of culture,washed three times, and resuspended at 1×10⁸ cells/mL in sterile saline.Groups of five naïve (unsensitized) C57BL/c mice were injected i.p. with200 μL of the cell suspension (2×10⁷ cells). Three days after transferof cells, the recipient mice were challenged with a 12-minute inhalationof an aerosol formulation of 1.5% OVA in saline (weight/volume), themist being produced by a nebulizer (small particle generator modelSPAG-2, ICN Pharmaceuticals, Costa Mesa, Calif.). Three days afteraerosol challenge, the mice were anesthetized with an i.p. injection ofan anesthetic mixture comprising Ketamine, acepromazine, and xylazine.The trachea of each mouse was exposed and cannulated. The lungs andupper airways were lavaged with 0.5 mL of cold (5° C.) phosphatebuffered saline (PBS). The cells within a 200 μL portion of thebronchoalveolar lavage (BAL) fluid were enumerated using a Coultercounter (Model ZB 1, Coulter Electronics, Hialeah, Fla.). The remainingBAL fluid was then centrifuged at 300×g for 5 minutes, and the cellsresuspended in 1 mL of Hank's balanced salt solution (HBSS, Gibco BRL),containing 0.5% fetal calf serum, and 10 mM ofN-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (HEPES, Gibco BRL).The cell suspension (100 μL) was centrifuged in a cytospin (ShandonSouthern Instruments, Sewickley, Pa.) and stained with Diff Quick todistinguish neutrophil, eosinophil, monocyte, and lymphocyte subsets.The number of eosinophils in the BAL fluid was determined by multiplyingthe percentage of eosinophils by the total cell count.

[0406] As shown in Table 2, OVA-sensitized splenocytes cultured in theabsence of MEK inhibitor, when transferred to naïve recipient mice, wereable to promote eosinophilic lung inflammation in response to an aerosolchallenge with OVA. In contrast, splenocytes cultured in the presence ofthe MEK inhibitors PD 171984, PD 184352, and PD 184386 (10 μM each) didnot promote eosinophilic lung inflammation (>99% inhibition). For eachof the MEK inhibitors used, a 10 μM concentration was previously foundto inhibit IL-5 production by over 75% (Table 1). These results suggestthat the MEK inhibitors inhibit the IL-5-producing T lymphocytes thatare required to support asthma-like eosinophilic lung inflammation inmice. TABLE 2 The Effects of MEK Inhibitors on the Adoptive-Transfer ofEosinophilic Lung Inflammation Treatment of Spleen Cell Culture %Inhibition of Compound Dose (μM) BAL Eosinophils None 0 PD 171984 1099.82 PD 184386 10 99.78 PD 184352 10 99.46

[0407] The final set of experiments was designed to test whether MEKinhibitors could inhibit active OVA-induced eosinophilic lunginflammation in mice. Mice were sensitized with OVA/aluminum hydroxideon Day 0 as described above. On Day 14, the mice were challenged byaerosol with 1.5% OVA, as described above for the adoptive-transferrecipients. One group of eight sensitized mice was dosed orally withvehicle (0.5% hydroxypropylmethylcellulose/0.25% TWEEN-80). Other groupsof sensitized mice (8 mice per group) were given oral doses of a MEKinhibitor. The test compound was dissolved in the vehicle, and thevolume for each dosage was adjusted to 200 μL, so that each test animalreceived the same oral volume. In experiments reported in Table 3 andTable 4, the MEK inhibitor was administered starting on Day 13 (ie, 13days after initial sensitization and 1 day prior to aerosol challenge),and continued daily through Day 16 (4 days total). In experimentsreported in Table 5, the MEK inhibitor was administered starting on Day7 (ie, 7 days after initial sensitization and 7 days prior to aerosolchallenge), and continued daily through Day 16 (9 days total). On Day 17of each experiment (17 days following the initial OVA challenge, and 3days after the OVA aerosol challenge), all animals including controlswere anesthetized, cannulated, and ravaged as previously described. Thenumber of BAL eosinophils was determined as described above.

[0408] In the initial analysis of active OVA-induced lung inflammation,multiple MEK inhibitors (PD 171984, PD 177168, PD 184161, PD 184386, andPD 184352) were dosed orally for 4 days. Only one compound, PD 171984,demonstrated any inhibition of pulmonary eosinophilia (Table 3). PD171984, along with PD 184352, were tested again at multiple doses, againdosing for only 4 days. The results in Table 4 essentially parallelthose in Table 3 for these compounds; PD 171984 appears active, whereasPD 184352 does not. As reported in Table 5, increasing the oral dosingschedule from 4 days to 9 days (7 days prior to aerosol challenge, 2days after) resulted in a degree of inhibitory activity for PD 184352 at100 mg/kg (59.85% inhibition, p=0.11). PD 171984 continued todemonstrate statistically significant inhibitory activity under thisdosing regimen.

[0409] In total, these results indicate that MEK inhibitors, when usedin vitro, are potent inhibitors of IL-5 production, and completelyinhibit the ability of antigen-stimulated cells to adoptively transferasthma-like symptoms to naïve recipient mice. When used in vivo, someMEK inhibitors are more active than others. However, a less potentcompound (PD 184352) was shown to inhibit the asthma-like response inmice under a more rigorous dosing regimen. Thus, the foregoing dataestablish that the selective MEK inhibitors are active in inhibiting amodel of asthma in mice. The compounds have little or no toxic effects,and accordingly are particularly well-suited for treating andcontrolling asthma in children, as well as adults. The compounds will beformulated for convenient oral or parenteral administration, includingby aerosol delivery, transdermal delivery, or even suppositories, andwill be administered in an antiasthmatic effective dose, which is thatamount that is effective to treat the particular asthma severity forwhich treatment is needed or otherwise desired. TABLE 3 The Effect ofMEK Inhibitors on Eosinophilic Lung Inflammation in Mice MEK InhibitorDose (μM) % Inhibition of BAL Eosinophilia PD 171984 100 55.26* PD177168 100 −123.38 PD 184161 100 −32.53 PD 184386 100 −33.24 PD 184352150 −5.41

[0410] TABLE 4 Inhibition of BAL Eosinophils With 4-Day Compound Dosing% Inhibition of BAL Eosinophils by: Dose (mg/kg) PD 184352 PD 171984 0 00 10 −22.7 38.4* 30 −153.8 46.6* 100 −8.2 58.4*

[0411] TABLE 5 Inhibition of BAL Eosinophils With 9-Day Compound Dosing% Inhibition of BAL Eosinophils by: PD 171984 Dose (mg/kg) PD 184352Experiment 1 Experiment 2 0 0 0 0 10 −42.99 −13.74 50.15* 30 1.83 80.64*37.23* 100 59.85 88.40* 54.77*

[0412] The foregoing data establish that the selective MEK inhibitorsare active in both inhibiting and controlling the asthmatic disease, forexample, prior to actual challenge and following challenge. Thecompounds are therefore useful in the prophylaxis of asthma, and also intreating and alleviating the symptoms that accompany the disease duringits active stage. The compounds have little or no toxic effects, andaccordingly are particularly well-suited for treating and controllingasthma in children, as well as adults. The compounds will be formulatedfor convenient oral or parenteral administration, including by aerosoldelivery, transdermal delivery, or even suppositories, and will beadministered in an antiasthmatic effective dose, which is that amountthat is effective to treat the particular asthma severity for whichtreatment is needed or otherwise desired.

[0413] D. Other Embodiments

[0414] From the above disclosure and examples, and from the claimsbelow, the essential features of the invention are readily apparent. Thescope of the invention also encompasses various modifications andadaptations within the knowledge of a person of ordinary skill. Examplesinclude a disclosed compound modified by addition or removal of aprotecting group, or an ester, pharmaceutical salt, hydrate, acid, oramide of a disclosed compound. Publications cited herein are herebyincorporated by reference in their entirety.

What is claimed is:
 1. A method for preventing or treating asthma inpatients, said method comprising the step of administering anantiasthmatic-effective amount of a MEK inhibitor to a patient in needof treatment, or to a patient suspected of developing asthma and in needof prophylactic treatment.
 2. A method according to claim 1 wherein thepatient being treated has been diagnosed as having asthma and is in needof treatment.
 3. A method according to claim 1, wherein the MEKinhibitor is a selective MEK inhibitor.